For pancreatic islet transplantation, one of the most important steps of islet isolation is islet purification. The most common method of islet purification is density gradient centrifugation because there are differences in density between islets and acinar tissue. However, the density of islets/acinar tissue depends on several conditions, such as the incubation time before purification and the osmolality of the preincubation solution. In this study, we evaluated the impact of using two different preincubation solutions before purification. We used the University of Wisconsin (UW) solution and a new preservation solution (HN-1), which we recently developed. There were no significant differences between the two solutions in terms of the islet yield, rate of viability, and purity or stimulation index after purification. There were also no differences in the attainability and suitability of posttransplantation normoglycemia. Our study shows that the HN-1 solution is equivalent to the UW solution for preincubation before islet purification.
{"title":"Comparison of Incubation Solutions Prior to the Purification of Porcine Islet Cells.","authors":"Takashi Kawai, Hirofumi Noguchi, Takashi Kuise, Atsuko Nakatsuka, Akihiro Katayama, Noriko Imagawa, Hitomi Usui Kataoka, Issei Saitoh, Yasufumi Noguchi, Masami Watanabe, Toshiyoshi Fujiwara","doi":"10.3727/215517913X674180","DOIUrl":"10.3727/215517913X674180","url":null,"abstract":"<p><p>For pancreatic islet transplantation, one of the most important steps of islet isolation is islet purification. The most common method of islet purification is density gradient centrifugation because there are differences in density between islets and acinar tissue. However, the density of islets/acinar tissue depends on several conditions, such as the incubation time before purification and the osmolality of the preincubation solution. In this study, we evaluated the impact of using two different preincubation solutions before purification. We used the University of Wisconsin (UW) solution and a new preservation solution (HN-1), which we recently developed. There were no significant differences between the two solutions in terms of the islet yield, rate of viability, and purity or stimulation index after purification. There were also no differences in the attainability and suitability of posttransplantation normoglycemia. Our study shows that the HN-1 solution is equivalent to the UW solution for preincubation before islet purification. </p>","PeriodicalId":9780,"journal":{"name":"Cell medicine","volume":"6 1-2 1","pages":"9-14"},"PeriodicalIF":0.0,"publicationDate":"2013-10-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3727/215517913X674180","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69755965","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2013-10-01DOI: 10.3727/215517913X674162
H. Noguchi
On behalf of the Japan Society for Organ Preservation and Medical Biology (JSOPMB), I express my sincere appreciation to Professor Paul R. Sanberg (Department of Neurosurgery and Brain Repair, Morsani College of Medicine, University of South Florida, FL, USA), Executive Editor of Cell Medicine, for providing us such an excellent opportunity to publish the data that were presented at the annual meeting of the JSOPMB. I also thank Dr. David Eve, Associate Editor of Cell Medicine, for the editing of our articles in detail. I am very sure that the relationship between Cell Medicine and JSOPMB has enhanced the motivation of JSOPMB members as well as board members and will continue to do so in the future, while also encouraging young Japanese researchers to join this organization. One of the extremely important missions of the annual meeting of the JSOPMB is to exchange new research outcomes and create new therapeutic concepts. JSOPMB always encourages and motivates young investigators. JSOPMB was started in 1974 for the study of organ preservation and developed widely in the 1990s with the participation of researchers in various fields of medicine, pharmacology, engineering, veterinary medicine, and basic science. Currently, JSOPMB has more than 400 members and is run under the direction of Dr. Takehide Asano, the president of the JSOPMB. Excellent presentations conducted at the 39th annual meeting of the JSOPMB held November 16–17, 2012, in Fukushima, Japan, under the supervision of Dr. Mitsukazu Gotoh (Professor, Department of Regenerative Surgery, Fukushima Medical University, Fukushima, Japan), were selected and given an opportunity to be published in this special issue of Cell Medicine. Twelve of these presentations are herein published in this special JSOPMB issue. There were five articles regarding pancreatic islet transplantation. Katayama et al. compared University of Wisconsin (UW) solution (gold standard preservation solution) with new preservation solution, HN-1, in pancreas preservation for islet isolation. Islet yield and islet graft function were significantly higher in the HN-1 group than the UW group. Kawai et al. evaluated the impact of UW and HN-1 solutions for preincubation before purification. The efficacy of the preincubation solutions and the quality of the isolated islets were similar. Kubota et al. studied the viability of Rho-associated protein kinase (ROCK) inhibitor Y-27632 in a culture system in vitro on isolated islets. Y-27632 inhibited cell apoptosis in a graft and was also indicated as effective in insulin secretion. Seita et al. created type 1 diabetes canine models that were suitable for the assessment of cell therapies, such as islet transplantation and bioartificial pancreas, with low-dose streptozotocin (STZ) injection and partial pancreatectomy. Yamashita et al. explored the potential of internationally transporting human islets from Alberta, Canada to Tokyo, Japan and obtaining viable dispersed islet ce
我谨代表日本器官保存与医学生物学学会(JSOPMB)向《细胞医学》杂志的执行编辑Paul R. Sanberg教授(美国佛罗里达州南佛罗里达大学莫尔萨尼医学院神经外科与脑修复系)表示衷心的感谢，感谢他为我们提供了这样一个绝佳的机会来发表在JSOPMB年会上提交的数据。我还要感谢《细胞医学》副主编David Eve博士对我们文章的详细编辑。我非常确信，细胞医学与JSOPMB之间的关系增强了JSOPMB成员和董事会成员的动力，并将在未来继续这样做，同时也鼓励年轻的日本研究人员加入这个组织。JSOPMB年会的一个极其重要的任务是交流新的研究成果和创造新的治疗理念。JSOPMB一直鼓励和激励年轻的调查人员。JSOPMB成立于1974年，目的是研究器官保存，在20世纪90年代得到了医学、药理学、工程、兽医学和基础科学等各个领域的研究人员的广泛发展。目前，JSOPMB有400多名成员，由JSOPMB会长浅野武雄博士指导。2012年11月16日至17日在日本福岛举行的JSOPMB第39届年会上，在Mitsukazu Gotoh博士(日本福岛医科大学再生外科教授)的监督下，优秀的报告被选中并有机会发表在这期《细胞医学》特刊上。其中的12个演讲将在本期JSOPMB特刊中发表。有5篇关于胰岛移植的文章。Katayama等人比较了威斯康星大学(University of Wisconsin, UW)溶液(金标准保存液)与新型保存液HN-1在胰岛分离胰腺保存中的应用。HN-1组胰岛产量和胰岛移植功能显著高于UW组。Kawai等人评估了UW和HN-1溶液在纯化前进行预孵育的影响。预孵育液的效果和离体胰岛质量相似。Kubota等人研究了rho相关蛋白激酶(ROCK)抑制剂Y-27632在离体胰岛体外培养系统中的生存能力。Y-27632抑制移植物细胞凋亡，并对胰岛素分泌有效。Seita等人创建的1型糖尿病犬模型适合于评估细胞疗法，如胰岛移植和生物人工胰腺，低剂量链脲佐菌素(STZ)注射和部分胰腺切除术。Yamashita等人探索了将人类胰岛从加拿大阿尔伯塔省国际运输到日本东京并获得可存活的分散胰岛细胞的潜力。有两篇关于移植免疫抑制的文章。Sasahara等人开发了一种新的检测方法，用CellTiter-Glo检测免疫抑制药物的敏感性，该方法测量细胞ATP的数量，以帮助提高检测方法的敏感性。用CellTiter-Glo法进行淋巴细胞免疫抑制剂敏感性试验(LIST)所需的血量减少到以前用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基溴化四唑(MTT)法进行LIST所需血量的25%。Sugiyama等人通过LIST评估了肾移植前后霉酚酸(MPA)的药理学疗效，并将其与这些受体的临床结果进行了比较。尽管在移植前后评估的MPA药理疗效与排斥事件发生率无关，但在肾移植后2周，低MPA药理疗效组的巨细胞病毒(CMV)感染发生率明显高于高MPA敏感性组。有两篇关于肝脏/肝细胞的文章。Hsu等人建立了一种由新生儿STZ注射和高脂肪饮食诱导的大鼠非酒精性脂肪性肝炎(NASH)模型，该模型几乎模仿了人类NASH的所有特征。Enosawa等人在大鼠模型中研究了抗氧化电子治疗对2型糖尿病诱导的NASH的疗效。在NASH大鼠模型中，极低能量的抗氧化电子治疗可减弱致病性升高的肝脏炎症和氧化应激，同时可能会损害葡萄糖代谢。干细胞研究是一个重要的话题。有三篇关于干细胞的文章。村上等人。 研究表明，Sandos近系小鼠(SIM)衍生的6-硫鸟嘌呤和瓦阿巴因耐药(STO)细胞可以吞噬变形链球菌(引起蛀牙的细菌)，这些细胞经常被用于建立胚胎干细胞/诱导多能干细胞(ES/iPS)并使其保持未分化状态。这些细菌总是污染主要分离的人类乳牙髓细胞的培养物，因此在使用这些细胞时是一个重要的考虑因素。Yukawa等研究表明，量子点(QDs)与传统探针相比具有明显的荧光能力，可用于iPS细胞的荧光标记。Miyazaki等人开发了用于脂肪组织源性干细胞成像的无镉量子点。这期JSOPMB的主题是“器官生物学的新发展”。董事会成员和我都期待着JSOPMB与细胞医学联合取得进一步进展。
{"title":"Organ Biology-New Development.","authors":"H. Noguchi","doi":"10.3727/215517913X674162","DOIUrl":"https://doi.org/10.3727/215517913X674162","url":null,"abstract":"On behalf of the Japan Society for Organ Preservation and Medical Biology (JSOPMB), I express my sincere appreciation to Professor Paul R. Sanberg (Department of Neurosurgery and Brain Repair, Morsani College of Medicine, University of South Florida, FL, USA), Executive Editor of Cell Medicine, for providing us such an excellent opportunity to publish the data that were presented at the annual meeting of the JSOPMB. I also thank Dr. David Eve, Associate Editor of Cell Medicine, for the editing of our articles in detail. I am very sure that the relationship between Cell Medicine and JSOPMB has enhanced the motivation of JSOPMB members as well as board members and will continue to do so in the future, while also encouraging young Japanese researchers to join this organization. \u0000 \u0000One of the extremely important missions of the annual meeting of the JSOPMB is to exchange new research outcomes and create new therapeutic concepts. JSOPMB always encourages and motivates young investigators. JSOPMB was started in 1974 for the study of organ preservation and developed widely in the 1990s with the participation of researchers in various fields of medicine, pharmacology, engineering, veterinary medicine, and basic science. Currently, JSOPMB has more than 400 members and is run under the direction of Dr. Takehide Asano, the president of the JSOPMB. \u0000 \u0000Excellent presentations conducted at the 39th annual meeting of the JSOPMB held November 16–17, 2012, in Fukushima, Japan, under the supervision of Dr. Mitsukazu Gotoh (Professor, Department of Regenerative Surgery, Fukushima Medical University, Fukushima, Japan), were selected and given an opportunity to be published in this special issue of Cell Medicine. Twelve of these presentations are herein published in this special JSOPMB issue. \u0000 \u0000There were five articles regarding pancreatic islet transplantation. Katayama et al. compared University of Wisconsin (UW) solution (gold standard preservation solution) with new preservation solution, HN-1, in pancreas preservation for islet isolation. Islet yield and islet graft function were significantly higher in the HN-1 group than the UW group. Kawai et al. evaluated the impact of UW and HN-1 solutions for preincubation before purification. The efficacy of the preincubation solutions and the quality of the isolated islets were similar. Kubota et al. studied the viability of Rho-associated protein kinase (ROCK) inhibitor Y-27632 in a culture system in vitro on isolated islets. Y-27632 inhibited cell apoptosis in a graft and was also indicated as effective in insulin secretion. Seita et al. created type 1 diabetes canine models that were suitable for the assessment of cell therapies, such as islet transplantation and bioartificial pancreas, with low-dose streptozotocin (STZ) injection and partial pancreatectomy. Yamashita et al. explored the potential of internationally transporting human islets from Alberta, Canada to Tokyo, Japan and obtaining viable dispersed islet ce","PeriodicalId":9780,"journal":{"name":"Cell medicine","volume":"6 1-2 1","pages":"1-2"},"PeriodicalIF":0.0,"publicationDate":"2013-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3727/215517913X674162","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69755853","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2013-08-29DOI: 10.3727/215517913X666549
T. Teratani, E. Kobayashi
Research in the life sciences has been greatly advanced by the ability to directly visualize cells, tissues, and organs. Preclinical studies often involve many small and large animal experiments and, frequently, cell and organ transplantations. The rat is an excellent animal model for the development of transplantation and surgical techniques because of its small size and ability to breed in small spaces. Ten years ago, we established color-imaging transgenic rats and methods for the direct visualization of their tissues. Since then, our transgenic rats have been used throughout the various fields that are concerned with cell transplantation therapy. In this minireview, we summarize results from some of the groups that have used our transgenic rats at the bench level and in cell transplantation research.
{"title":"Bioimaging of Transgenic Rats Established at Jichi Medical University: Applications in Transplantation Research.","authors":"T. Teratani, E. Kobayashi","doi":"10.3727/215517913X666549","DOIUrl":"https://doi.org/10.3727/215517913X666549","url":null,"abstract":"Research in the life sciences has been greatly advanced by the ability to directly visualize cells, tissues, and organs. Preclinical studies often involve many small and large animal experiments and, frequently, cell and organ transplantations. The rat is an excellent animal model for the development of transplantation and surgical techniques because of its small size and ability to breed in small spaces. Ten years ago, we established color-imaging transgenic rats and methods for the direct visualization of their tissues. Since then, our transgenic rats have been used throughout the various fields that are concerned with cell transplantation therapy. In this minireview, we summarize results from some of the groups that have used our transgenic rats at the bench level and in cell transplantation research.","PeriodicalId":9780,"journal":{"name":"Cell medicine","volume":"5 2-3 1","pages":"45-51"},"PeriodicalIF":0.0,"publicationDate":"2013-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3727/215517913X666549","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69755884","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2013-08-10DOI: 10.3727/215517913X666468
W. Wolber, Ruhel Ahmad, S. Choi, S. Eckardt, K. McLaughlin, Jessica Schmitt, C. Geis, M. Heckmann, A. Sirén, A. Müller
Uniparental zygotes with two paternal (androgenetic, AG) or two maternal genomes (gynogenetic, GG) cannot develop into viable offsprings but form blastocysts from which pluripotent embryonic stem (ES) cells can be derived. For most organs, it is unclear whether uniparental ES cells can give rise to stably expandable somatic stem cells that can repair injured tissues. Even if previous reports indicated that the capacity of AG ES cells to differentiate in vitro into pan-neural progenitor cells (pNPCs) and into cells expressing neural markers is similar to biparental [normal fertilized (N)] ES cells, their potential for functional neurogenesis is not known. Here we show that murine AG pNPCs give rise to neuron-like cells, which then generate sodium-driven action potentials while maintaining fidelity of imprinted gene expression. Neural engraftment after intracerebral transplantation was achieved only by late (22 days) AG and N pNPCs with in vitro low colony-forming cell (CFC) capacity. However, persisting CFC formation seen, in particular, in early (13 or 16 days) differentiation cultures of N and AG pNPCs correlated with a high incidence of trigerm layer teratomas. As AG ES cells display functional neurogenesis and in vivo stability similar to N ES cells, they represent a unique model system to study the roles of paternal and maternal genomes on neural development and on the development of imprinting-associated brain diseases.
{"title":"Phenotype and Stability of Neural Differentiation of Androgenetic Murine ES Cell-Derived Neural Progenitor Cells.","authors":"W. Wolber, Ruhel Ahmad, S. Choi, S. Eckardt, K. McLaughlin, Jessica Schmitt, C. Geis, M. Heckmann, A. Sirén, A. Müller","doi":"10.3727/215517913X666468","DOIUrl":"https://doi.org/10.3727/215517913X666468","url":null,"abstract":"Uniparental zygotes with two paternal (androgenetic, AG) or two maternal genomes (gynogenetic, GG) cannot develop into viable offsprings but form blastocysts from which pluripotent embryonic stem (ES) cells can be derived. For most organs, it is unclear whether uniparental ES cells can give rise to stably expandable somatic stem cells that can repair injured tissues. Even if previous reports indicated that the capacity of AG ES cells to differentiate in vitro into pan-neural progenitor cells (pNPCs) and into cells expressing neural markers is similar to biparental [normal fertilized (N)] ES cells, their potential for functional neurogenesis is not known. Here we show that murine AG pNPCs give rise to neuron-like cells, which then generate sodium-driven action potentials while maintaining fidelity of imprinted gene expression. Neural engraftment after intracerebral transplantation was achieved only by late (22 days) AG and N pNPCs with in vitro low colony-forming cell (CFC) capacity. However, persisting CFC formation seen, in particular, in early (13 or 16 days) differentiation cultures of N and AG pNPCs correlated with a high incidence of trigerm layer teratomas. As AG ES cells display functional neurogenesis and in vivo stability similar to N ES cells, they represent a unique model system to study the roles of paternal and maternal genomes on neural development and on the development of imprinting-associated brain diseases.","PeriodicalId":9780,"journal":{"name":"Cell medicine","volume":"5 1 1","pages":"29-42"},"PeriodicalIF":0.0,"publicationDate":"2013-08-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3727/215517913X666468","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69756052","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2013-08-01DOI: 10.3727/215517913X666521
H. Hsu, N. Matsuno, N. Machida, S. Enosawa
Demand for human primary hepatocytes is increasing, particularly for clinical trials of hepatocyte transplantation. However, due to the severe shortage of organ transplant donors, the source of cells for these endeavors is restricted to untransplantable livers, such as those from non-heart-beating donors and surgically resected liver tissues. To improve cell recovery from such sources after warm ischemia, we evaluated the efficacy of applying perfusion solutions, focusing on improvement of hepatocyte recovery. Warm ischemia was induced by clamping both portal vein and hepatic artery for 10 or 15 min in rats. The liver was perfused with either Euro-Collins (EC) or extracellular-type trehalose-containing Kyoto (ETK) solutions supplemented with an anticoagulant, either heparin or citrate phosphate dextrose solution (CPD), compared to Ca(2+), Mg(2+)-free Hanks solution. While the viability of recovered cells was 81.5 ± 4.2% and cell yield was 2.27 ± 0.53 × 10(8) in nonwarm ischemia controls (n = 11), these values were only 74.7 ± 2.9% and 0.38 ± 0.17 × 10(8), respectively, in the 10-min warm ischemia group, using the Hanks as the perfusion solution. Although the addition of heparin increased the live cell number only twofold (0.71 ± 0.40 × 10(8), n = 4), the best improvement was achieved by adding CPD to EC. This resulted in a recovery of 1.41 ± 0.50 × 10(8) in the 10-min ischemia group (n = 7) and 1.37 ± 0.28 × 10(8) in the 15-min group (n = 3). Macroscopic observation showed that blood had been completely flushed out by the solution, suggesting good restoration of the microcirculation in ischemic liver. Using ETK instead of EC resulted in a slight decrease in efficacy. These results demonstrate that CPD, as opposed to heparin, is effective in ensuring liver microcirculation and flushing out the blood and that EC is the best perfusion solution for obtaining hepatocytes from ischemic liver.
{"title":"Improved Recovery of Hepatocytes Isolated From Warm Ischemic Rat Liver by Citrate Phosphate Dextrose (CPD)-Supplemented Euro-Collins Solution.","authors":"H. Hsu, N. Matsuno, N. Machida, S. Enosawa","doi":"10.3727/215517913X666521","DOIUrl":"https://doi.org/10.3727/215517913X666521","url":null,"abstract":"Demand for human primary hepatocytes is increasing, particularly for clinical trials of hepatocyte transplantation. However, due to the severe shortage of organ transplant donors, the source of cells for these endeavors is restricted to untransplantable livers, such as those from non-heart-beating donors and surgically resected liver tissues. To improve cell recovery from such sources after warm ischemia, we evaluated the efficacy of applying perfusion solutions, focusing on improvement of hepatocyte recovery. Warm ischemia was induced by clamping both portal vein and hepatic artery for 10 or 15 min in rats. The liver was perfused with either Euro-Collins (EC) or extracellular-type trehalose-containing Kyoto (ETK) solutions supplemented with an anticoagulant, either heparin or citrate phosphate dextrose solution (CPD), compared to Ca(2+), Mg(2+)-free Hanks solution. While the viability of recovered cells was 81.5 ± 4.2% and cell yield was 2.27 ± 0.53 × 10(8) in nonwarm ischemia controls (n = 11), these values were only 74.7 ± 2.9% and 0.38 ± 0.17 × 10(8), respectively, in the 10-min warm ischemia group, using the Hanks as the perfusion solution. Although the addition of heparin increased the live cell number only twofold (0.71 ± 0.40 × 10(8), n = 4), the best improvement was achieved by adding CPD to EC. This resulted in a recovery of 1.41 ± 0.50 × 10(8) in the 10-min ischemia group (n = 7) and 1.37 ± 0.28 × 10(8) in the 15-min group (n = 3). Macroscopic observation showed that blood had been completely flushed out by the solution, suggesting good restoration of the microcirculation in ischemic liver. Using ETK instead of EC resulted in a slight decrease in efficacy. These results demonstrate that CPD, as opposed to heparin, is effective in ensuring liver microcirculation and flushing out the blood and that EC is the best perfusion solution for obtaining hepatocytes from ischemic liver.","PeriodicalId":9780,"journal":{"name":"Cell medicine","volume":"5 2-3 1","pages":"97-101"},"PeriodicalIF":0.0,"publicationDate":"2013-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3727/215517913X666521","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69755763","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2013-05-14DOI: 10.3727/215517913X666558
M. Maruyama, T. Kenmochi, N. Akutsu, K. Otsuki, T. Ito, I. Matsumoto, T. Asano
Autologous islet transplantation after total or semitotal pancreatectomy aims to preserve insulin secretory function and prevent the onset of diabetes. The major indication for pancreatectomy is chronic pancreatitis with severe abdominal pain, a benign pancreatic tumor, and trauma. The metabolic outcome of autologous islet transplantation is better than that of allogeneic transplantation and depends on the number of transplanted islets. Achieving islet isolation from a fibrous or damaged pancreas is one of the biggest challenges of autologous islet transplantation; a major complication is portal vein thrombosis after crude islet infusion. However, the incidence of portal vein thrombosis has decreased as islet preparation techniques have improved over time.
{"title":"A Review of Autologous Islet Transplantation.","authors":"M. Maruyama, T. Kenmochi, N. Akutsu, K. Otsuki, T. Ito, I. Matsumoto, T. Asano","doi":"10.3727/215517913X666558","DOIUrl":"https://doi.org/10.3727/215517913X666558","url":null,"abstract":"Autologous islet transplantation after total or semitotal pancreatectomy aims to preserve insulin secretory function and prevent the onset of diabetes. The major indication for pancreatectomy is chronic pancreatitis with severe abdominal pain, a benign pancreatic tumor, and trauma. The metabolic outcome of autologous islet transplantation is better than that of allogeneic transplantation and depends on the number of transplanted islets. Achieving islet isolation from a fibrous or damaged pancreas is one of the biggest challenges of autologous islet transplantation; a major complication is portal vein thrombosis after crude islet infusion. However, the incidence of portal vein thrombosis has decreased as islet preparation techniques have improved over time.","PeriodicalId":9780,"journal":{"name":"Cell medicine","volume":"5 2-3 1","pages":"59-62"},"PeriodicalIF":0.0,"publicationDate":"2013-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3727/215517913X666558","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69755898","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2013-05-14DOI: 10.3727/215517913X666495
H. Noguchi, I. Saitoh, H. Kataoka, Masami Watanabe, Yasufumi Noguchi, T. Fujiwara
Recently, mouse pancreatic stem cells have been isolated from adult mouse pancreata. However, these pancreatic stem cells could be maintained only under specific culture conditions with lot-limited fetal bovine serum (FBS). For the efficient isolation and maintenance of mouse pancreatic stem cells, it is important to identify culture conditions that can be used independent of the FBS lot. In this study, we evaluated the culture conditions required to maintain mouse pancreatic stem cells. The mouse pancreatic stem cells derived from the pancreas of a newborn mouse, HN#101, were cultured under the following conditions: 1) Dulbecco's modified Eagle's medium (DMEM) with 20% lot-limited FBS, in which mouse pancreatic stem cells could be cultured without changes in morphology and growth activity; 2) complete embryonic stem (ES) cell media; and 3) complete ES cell media on feeder layers of mitomycin C-treated STO cells, which were the same culture conditions used for mouse ES cells. Under culture conditions #1 and #3, the HN#101 cells continued to form a flat "cobblestone" monolayer and continued to divide actively beyond the population doubling level (PDL) 100 without growth inhibition, but this did not occur under culture condition #2. The gene expression profile and differentiated capacity of the HN#101 cells cultured for 2 months under culture condition #3 were similar to those of HN#101 cells at PDL 50. These data suggest that complete ES cell media on feeder layers could be useful for maintaining the undifferentiated state of pancreatic stem cells.
近年来，从成年小鼠胰腺中分离出小鼠胰腺干细胞。然而，这些胰腺干细胞只能在特定的培养条件下与lot-limited胎牛血清(FBS)维持。为了有效地分离和维持小鼠胰腺干细胞，确定可以独立于FBS批次使用的培养条件是很重要的。在这项研究中，我们评估了维持小鼠胰腺干细胞所需的培养条件。采用新生小鼠HN#101胰腺提取的小鼠胰腺干细胞，在以下条件下进行培养:1)Dulbecco's modified Eagle's medium (DMEM)中添加20%的批限FBS，在此条件下，小鼠胰腺干细胞可以在无形态学和生长活性变化的情况下培养;2)完全胚胎干细胞培养基;3)在丝裂霉素c处理的STO细胞的饲养层上添加完整的ES细胞培养基，与小鼠ES细胞的培养条件相同。在培养条件#1和#3下，hn# 101细胞继续形成扁平的“鹅卵石”单层，并在超过种群倍增水平(PDL) 100后继续积极分裂，没有生长抑制，但在培养条件#2下没有发生这种情况。在培养条件3下培养2个月的hn# 101细胞的基因表达谱和分化能力与PDL 50下的hn# 101细胞相似。这些数据表明，饲喂层上完整的胚胎干细胞培养基可能有助于维持胰腺干细胞的未分化状态。
{"title":"Culture Conditions for Mouse Pancreatic Stem Cells.","authors":"H. Noguchi, I. Saitoh, H. Kataoka, Masami Watanabe, Yasufumi Noguchi, T. Fujiwara","doi":"10.3727/215517913X666495","DOIUrl":"https://doi.org/10.3727/215517913X666495","url":null,"abstract":"Recently, mouse pancreatic stem cells have been isolated from adult mouse pancreata. However, these pancreatic stem cells could be maintained only under specific culture conditions with lot-limited fetal bovine serum (FBS). For the efficient isolation and maintenance of mouse pancreatic stem cells, it is important to identify culture conditions that can be used independent of the FBS lot. In this study, we evaluated the culture conditions required to maintain mouse pancreatic stem cells. The mouse pancreatic stem cells derived from the pancreas of a newborn mouse, HN#101, were cultured under the following conditions: 1) Dulbecco's modified Eagle's medium (DMEM) with 20% lot-limited FBS, in which mouse pancreatic stem cells could be cultured without changes in morphology and growth activity; 2) complete embryonic stem (ES) cell media; and 3) complete ES cell media on feeder layers of mitomycin C-treated STO cells, which were the same culture conditions used for mouse ES cells. Under culture conditions #1 and #3, the HN#101 cells continued to form a flat \"cobblestone\" monolayer and continued to divide actively beyond the population doubling level (PDL) 100 without growth inhibition, but this did not occur under culture condition #2. The gene expression profile and differentiated capacity of the HN#101 cells cultured for 2 months under culture condition #3 were similar to those of HN#101 cells at PDL 50. These data suggest that complete ES cell media on feeder layers could be useful for maintaining the undifferentiated state of pancreatic stem cells.","PeriodicalId":9780,"journal":{"name":"Cell medicine","volume":"5 2-3 1","pages":"63-8"},"PeriodicalIF":0.0,"publicationDate":"2013-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3727/215517913X666495","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69756129","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2013-05-14DOI: 10.3727/215517913X666477
N. Kasahara, T. Teratani, J. Doi, Yuki Iijima, M. Maeda, S. Uemoto, Y. Fujimoto, N. Sata, Y. Yasuda, E. Kobayashi
Pancreatic islet transplantation has received widespread attention as a promising treatment for type 1 diabetes. However, islets for transplantation are subject to damage from a number of sources, including ischemic injury during removal and delivery of the donor pancreas, enzymatic digestion during islet isolation, and reperfusion injury after transplantation in the recipient. Here we found that protein fractions secreted by mesenchymal stem cells (MSCs) were capable of activating preserved islets. A conditioned medium from the supernatant obtained by culturing adipose tissue MSCs (derived from wild-type Lewis rats) was prepared for 2 days in serum-free medium. Luc-Tg rat islets to which an organ preservation solution was added were then incubated at 4°C with fractions of various molecular weights prepared from the conditioned medium. Under the treatment with some of the fractions, by 4 days the relative luminescence intensities (representative of the ATP levels of the cold-preserved islets) had increased to over 150% of their initial values. Our novel system may be able to restore isolated islets to the condition they were in before transport, culture, and transplantation.
{"title":"Use of Mesenchymal Stem Cell-Conditioned Medium to Activate Islets in Preservation Solution.","authors":"N. Kasahara, T. Teratani, J. Doi, Yuki Iijima, M. Maeda, S. Uemoto, Y. Fujimoto, N. Sata, Y. Yasuda, E. Kobayashi","doi":"10.3727/215517913X666477","DOIUrl":"https://doi.org/10.3727/215517913X666477","url":null,"abstract":"Pancreatic islet transplantation has received widespread attention as a promising treatment for type 1 diabetes. However, islets for transplantation are subject to damage from a number of sources, including ischemic injury during removal and delivery of the donor pancreas, enzymatic digestion during islet isolation, and reperfusion injury after transplantation in the recipient. Here we found that protein fractions secreted by mesenchymal stem cells (MSCs) were capable of activating preserved islets. A conditioned medium from the supernatant obtained by culturing adipose tissue MSCs (derived from wild-type Lewis rats) was prepared for 2 days in serum-free medium. Luc-Tg rat islets to which an organ preservation solution was added were then incubated at 4°C with fractions of various molecular weights prepared from the conditioned medium. Under the treatment with some of the fractions, by 4 days the relative luminescence intensities (representative of the ATP levels of the cold-preserved islets) had increased to over 150% of their initial values. Our novel system may be able to restore isolated islets to the condition they were in before transport, culture, and transplantation.","PeriodicalId":9780,"journal":{"name":"Cell medicine","volume":"5 2-3 1","pages":"75-81"},"PeriodicalIF":0.0,"publicationDate":"2013-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3727/215517913X666477","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69756087","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2013-05-14DOI: 10.3727/215517913X666503
Takashi Kuise, H. Noguchi, I. Saitoh, H. Kataoka, Masami Watanabe, Yasufumi Noguchi, T. Fujiwara
Mouse pancreatic stem cells have been isolated from mouse pancreata. This study evaluated the efficacy of isolating mouse pancreatic stem cells using mice of different ages. The pancreata of newborn mice, 8-week-old mice, and 24-week-old mice were harvested and digested by using collagenase. The "duct-like" cells in the digested pancreatic tissue were then inoculated into 96-well plates, cloned by limiting dilution, and cultured in DMEM with 20% FBS. Pancreatic stem cells were isolated from the pancreata of all newborn mice, while cells could only be isolated from 10% of the pancreata of 8-week-old mice and could not be isolated from the pancreata of any 24-week-old mice. These data suggest that young mice may have some pancreatic stem cells and that older mice may only have a few pancreatic stem cells. These data also indicate that it is extremely difficult to isolate pancreatic stem cells from older mice, suggesting that future research focus its efforts on finding methods of isolating pancreatic stem cells from adult mice.
{"title":"Isolation Efficiency of Mouse Pancreatic Stem Cells Is Age Dependent.","authors":"Takashi Kuise, H. Noguchi, I. Saitoh, H. Kataoka, Masami Watanabe, Yasufumi Noguchi, T. Fujiwara","doi":"10.3727/215517913X666503","DOIUrl":"https://doi.org/10.3727/215517913X666503","url":null,"abstract":"Mouse pancreatic stem cells have been isolated from mouse pancreata. This study evaluated the efficacy of isolating mouse pancreatic stem cells using mice of different ages. The pancreata of newborn mice, 8-week-old mice, and 24-week-old mice were harvested and digested by using collagenase. The \"duct-like\" cells in the digested pancreatic tissue were then inoculated into 96-well plates, cloned by limiting dilution, and cultured in DMEM with 20% FBS. Pancreatic stem cells were isolated from the pancreata of all newborn mice, while cells could only be isolated from 10% of the pancreata of 8-week-old mice and could not be isolated from the pancreata of any 24-week-old mice. These data suggest that young mice may have some pancreatic stem cells and that older mice may only have a few pancreatic stem cells. These data also indicate that it is extremely difficult to isolate pancreatic stem cells from older mice, suggesting that future research focus its efforts on finding methods of isolating pancreatic stem cells from adult mice.","PeriodicalId":9780,"journal":{"name":"Cell medicine","volume":"5 2-3 1","pages":"69-73"},"PeriodicalIF":0.0,"publicationDate":"2013-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3727/215517913X666503","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69756137","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2013-05-14DOI: 10.3727/215517913X666530
Y. Miyamoto, Yumie Koshidaka, H. Noguchi, Koichi Oishi, Hiroaki Saito, H. Yukawa, N. Kaji, T. Ikeya, Satoshi Suzuki, Hisashi Iwata, Y. Baba, K. Murase, S. Hayashi
Magnetic resonance imaging (MRI) using magnetic nanoparticles has been used to diagnose vascular diseases as well as to monitor transplanted cells and tissues. In this study, we synthesized magnetic iron oxide nanoparticles (TMADM-03), electrically charged by the presence of a cationic end-group substitution of dextran, and observed these nanoparticles inside three-dimensional models of HepG2 spheroids, which mimic tissues. Patterned cell array glass disks were prepared to visualize the presence of TMADM-03 uptaken by HepG2 spheroids using transmission electron microscopy (TEM). The HepG2 cells (2 × 10(5) cells) were inoculated onto Cell-able™ 12-well plates. After 48 h of culture, the cells were incubated with 75 µg Fe/ml TMADM-03 in culture medium for 24 h. To investigate the cellular function of the HepG2 spheroids, the albumin secretion was evaluated by an ELISA. The albumin secretion after incubation for 24 h was reduced compared with the secretion prior to the addition of TMADM-03. TEM image samples were prepared in a planar direction or a vertical direction to the HepG2 spheroids on patterned cell array glass disks. The incorporation of TMADM-03 inside the HepG2 spheroids was confirmed. In addition, TMADM-03 could be observed in the deeper layers of the spheroids, and this was localized in the lysosomes. These data suggest that the novel magnetic iron oxide nanoparticles invade three-dimensional HepG2 spheroids.
{"title":"Observation of Positively Charged Magnetic Nanoparticles Inside HepG2 Spheroids Using Electron Microscopy.","authors":"Y. Miyamoto, Yumie Koshidaka, H. Noguchi, Koichi Oishi, Hiroaki Saito, H. Yukawa, N. Kaji, T. Ikeya, Satoshi Suzuki, Hisashi Iwata, Y. Baba, K. Murase, S. Hayashi","doi":"10.3727/215517913X666530","DOIUrl":"https://doi.org/10.3727/215517913X666530","url":null,"abstract":"Magnetic resonance imaging (MRI) using magnetic nanoparticles has been used to diagnose vascular diseases as well as to monitor transplanted cells and tissues. In this study, we synthesized magnetic iron oxide nanoparticles (TMADM-03), electrically charged by the presence of a cationic end-group substitution of dextran, and observed these nanoparticles inside three-dimensional models of HepG2 spheroids, which mimic tissues. Patterned cell array glass disks were prepared to visualize the presence of TMADM-03 uptaken by HepG2 spheroids using transmission electron microscopy (TEM). The HepG2 cells (2 × 10(5) cells) were inoculated onto Cell-able™ 12-well plates. After 48 h of culture, the cells were incubated with 75 µg Fe/ml TMADM-03 in culture medium for 24 h. To investigate the cellular function of the HepG2 spheroids, the albumin secretion was evaluated by an ELISA. The albumin secretion after incubation for 24 h was reduced compared with the secretion prior to the addition of TMADM-03. TEM image samples were prepared in a planar direction or a vertical direction to the HepG2 spheroids on patterned cell array glass disks. The incorporation of TMADM-03 inside the HepG2 spheroids was confirmed. In addition, TMADM-03 could be observed in the deeper layers of the spheroids, and this was localized in the lysosomes. These data suggest that the novel magnetic iron oxide nanoparticles invade three-dimensional HepG2 spheroids.","PeriodicalId":9780,"journal":{"name":"Cell medicine","volume":"5 2-3 1","pages":"89-96"},"PeriodicalIF":0.0,"publicationDate":"2013-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3727/215517913X666530","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69755778","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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