Pub Date : 2013-12-30DOI: 10.3727/215517913X674207
H. Sasahara, K. Sugiyama, M. Tsukaguchi, K. Isogai, A. Toyama, H. Satoh, K. Saitoh, Y. Nakagawa, Kota Takahashi, Sachiko Tanaka, K. Onda, T. Hirano
The lymphocyte immunosuppressant sensitivity test (LIST) using patient peripheral lymphocytes can predict the therapeutic efficacy of immunosuppressive drugs used in renal transplantation. We have evaluated the pharmacological efficacy of drugs by using the LIST with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, which measures the cellular mitochondrial activity. The LIST with the MTT assay requires a relatively large amount of blood. As such, we developed a new assay for examining drug sensitivity with a CellTiter-Glo assay, which measures the amount of cellular ATP to help increase the assay's sensitivity and reduce the amount of blood needed. Renal transplant recipients generally receive either cyclosporine or tacrolimus, in addition to mycophenolate mofetil and methylprednisolone, as an immunosuppressive therapy to prevent acute rejection. We evaluated the pharmacological efficacy of these immunosuppressive agents with both the MTT and CellTiter-Glo assays using the peripheral blood mononuclear cells of 21 healthy volunteers. Furthermore, we also examined the relationship between these immunosuppressive agents' pharmacological efficacy and the results of the MTT and CellTiter-Glo assays. The IC50 values for cyclosporine, tacrolimus, mycophenolic acid, and methylprednisolone were significantly correlated between the MTT and CellTiter-Glo assays. The amount of blood cells required for LIST with the CellTiter-Glo assay was able to be reduced to 25% of the amount required for the previously established LIST with the MTT assay procedure. We concluded from these observations that the LIST with the CellTiter-Glo assay should be used instead of the MTT assay for carrying out individualized immunosuppressive therapy in renal transplantation patients.
{"title":"Comparison of the Pharmacological Efficacies of Immunosuppressive Drugs Evaluated by the ATP Production and Mitochondrial Activity in Human Lymphocytes.","authors":"H. Sasahara, K. Sugiyama, M. Tsukaguchi, K. Isogai, A. Toyama, H. Satoh, K. Saitoh, Y. Nakagawa, Kota Takahashi, Sachiko Tanaka, K. Onda, T. Hirano","doi":"10.3727/215517913X674207","DOIUrl":"https://doi.org/10.3727/215517913X674207","url":null,"abstract":"The lymphocyte immunosuppressant sensitivity test (LIST) using patient peripheral lymphocytes can predict the therapeutic efficacy of immunosuppressive drugs used in renal transplantation. We have evaluated the pharmacological efficacy of drugs by using the LIST with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, which measures the cellular mitochondrial activity. The LIST with the MTT assay requires a relatively large amount of blood. As such, we developed a new assay for examining drug sensitivity with a CellTiter-Glo assay, which measures the amount of cellular ATP to help increase the assay's sensitivity and reduce the amount of blood needed. Renal transplant recipients generally receive either cyclosporine or tacrolimus, in addition to mycophenolate mofetil and methylprednisolone, as an immunosuppressive therapy to prevent acute rejection. We evaluated the pharmacological efficacy of these immunosuppressive agents with both the MTT and CellTiter-Glo assays using the peripheral blood mononuclear cells of 21 healthy volunteers. Furthermore, we also examined the relationship between these immunosuppressive agents' pharmacological efficacy and the results of the MTT and CellTiter-Glo assays. The IC50 values for cyclosporine, tacrolimus, mycophenolic acid, and methylprednisolone were significantly correlated between the MTT and CellTiter-Glo assays. The amount of blood cells required for LIST with the CellTiter-Glo assay was able to be reduced to 25% of the amount required for the previously established LIST with the MTT assay procedure. We concluded from these observations that the LIST with the CellTiter-Glo assay should be used instead of the MTT assay for carrying out individualized immunosuppressive therapy in renal transplantation patients.","PeriodicalId":9780,"journal":{"name":"Cell medicine","volume":"6 1-2 1","pages":"39-45"},"PeriodicalIF":0.0,"publicationDate":"2013-12-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3727/215517913X674207","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69756029","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 : 2013-12-30DOI: 10.3727/215517913X674270
H. Yukawa, Kaoru Suzuki, Yukiko Kano, Tatsuya Yamada, N. Kaji, Tetsuya Ishikawa, Y. Baba
Induced pluripotent stem (iPS) cells have received remarkable attention as the cell sources for clinical applications of regenerative medicine including stem cell therapy. Additionally, labeling technology is in high demand for tracing transplanted cells used in stem cell therapy. In this study, we used quantum dots (QDs), which have distinct fluorescence abilities in comparison with traditional probes, as the labeling materials and investigated whether iPS cells could be labeled with QDs with no cytotoxicity. iPS cells could not be labeled with QDs alone but required the use of cell-penetrating peptides such as octaarginine (R8). No significant cytotoxicity to iPS cells was confirmed by up to 8 nM QDs, and the iPS cells labeled with QDs maintained their undifferentiated state and pluripotency. These data suggest that QDs can be used for fluorescence labeling of iPS cells.
{"title":"Induced Pluripotent Stem Cell Labeling Using Quantum Dots.","authors":"H. Yukawa, Kaoru Suzuki, Yukiko Kano, Tatsuya Yamada, N. Kaji, Tetsuya Ishikawa, Y. Baba","doi":"10.3727/215517913X674270","DOIUrl":"https://doi.org/10.3727/215517913X674270","url":null,"abstract":"Induced pluripotent stem (iPS) cells have received remarkable attention as the cell sources for clinical applications of regenerative medicine including stem cell therapy. Additionally, labeling technology is in high demand for tracing transplanted cells used in stem cell therapy. In this study, we used quantum dots (QDs), which have distinct fluorescence abilities in comparison with traditional probes, as the labeling materials and investigated whether iPS cells could be labeled with QDs with no cytotoxicity. iPS cells could not be labeled with QDs alone but required the use of cell-penetrating peptides such as octaarginine (R8). No significant cytotoxicity to iPS cells was confirmed by up to 8 nM QDs, and the iPS cells labeled with QDs maintained their undifferentiated state and pluripotency. These data suggest that QDs can be used for fluorescence labeling of iPS cells.","PeriodicalId":9780,"journal":{"name":"Cell medicine","volume":"6 1-2 1","pages":"83-90"},"PeriodicalIF":0.0,"publicationDate":"2013-12-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3727/215517913X674270","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69756360","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 : 2013-12-30DOI: 10.3727/215517913X674289
M. Seita, H. Noguchi, Yasuhiro Kubota, Hironobu Kawamoto, S. Nakaji, N. Kobayashi, T. Fujiwara
We created canine models of type 1 diabetes 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. In our model, a 50% pancreatectomy was performed with general anesthesia, followed by systemic injection of 35 mg/kg STZ into a vein of the foreleg. Four weeks after the administration of STZ, the fasting blood glucose level of our model dogs was found to be over 200 mg/dl twice on different days, and we could not detect any canine insulin by the intravenous glucose tolerance test (IVGTT). We therefore diagnosed the dogs to have induced diabetes. Some studies have reported high-dose STZ to be very toxic for both the kidney and liver, and therefore a lower dose is desirable to induce diabetic models without any associated kidney or liver damage. We think that the combination of a partial pancreatectomy can thus make it possible to reduce the dose of STZ, and it is therefore useful for the creation of type 1 diabetes models. We believe that our model is a safe and reliable model for type 1 diabetes in canines to assess the efficacy of pancreas-targeted cell therapies.
{"title":"Development of Canine Models of Type 1 Diabetes With Partial Pancreatectomy and the Administration of Streptozotocin.","authors":"M. Seita, H. Noguchi, Yasuhiro Kubota, Hironobu Kawamoto, S. Nakaji, N. Kobayashi, T. Fujiwara","doi":"10.3727/215517913X674289","DOIUrl":"https://doi.org/10.3727/215517913X674289","url":null,"abstract":"We created canine models of type 1 diabetes 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. In our model, a 50% pancreatectomy was performed with general anesthesia, followed by systemic injection of 35 mg/kg STZ into a vein of the foreleg. Four weeks after the administration of STZ, the fasting blood glucose level of our model dogs was found to be over 200 mg/dl twice on different days, and we could not detect any canine insulin by the intravenous glucose tolerance test (IVGTT). We therefore diagnosed the dogs to have induced diabetes. Some studies have reported high-dose STZ to be very toxic for both the kidney and liver, and therefore a lower dose is desirable to induce diabetic models without any associated kidney or liver damage. We think that the combination of a partial pancreatectomy can thus make it possible to reduce the dose of STZ, and it is therefore useful for the creation of type 1 diabetes models. We believe that our model is a safe and reliable model for type 1 diabetes in canines to assess the efficacy of pancreas-targeted cell therapies.","PeriodicalId":9780,"journal":{"name":"Cell medicine","volume":"6 1-2 1","pages":"25-31"},"PeriodicalIF":0.0,"publicationDate":"2013-12-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3727/215517913X674289","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69756374","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 : 2013-12-30DOI: 10.3727/215517913X674234
Tomoya Murakami, I. Saitoh, E. Inada, Mie Kurosawa, Y. Iwase, H. Noguchi, Y. Terao, Y. Yamasaki, H. Hayasaki, Masahiro Sato
STO feeder cells, a line established from mouse SIM embryonic fibroblasts, have been frequently used for establishing embryonic stem cells and maintaining them in an undifferentiated state. There are some reports demonstrating that fibroblastic cells have the ability to phagocytose Gram-positive bacterium (e.g., streptococci and staphylococci). In this study, we examined the possibility that STO cells could phagocytose Streptococcus mutans (a bacteria causing tooth decay), which always contaminates cultures of primarily isolated human deciduous dental pulp cells (HDDPCs). Simple cultivation of the primary HDDPCs in the absence of STO cells allowed S. mutans to massively propagate in the medium, thus leading to an opaque medium. In contrast, there was no bacterial contamination in the cultures containing mitomycin C (MMC)-inactivated STO cells. Furthermore, STO cells indicated bacterial phagocytic activity under fluorescent microscopy with the dye pHrodo. Besides removal of contaminating bacteria, STO feeder cells allowed the HDDPCs to spread out. These data suggest that MMC-treated STO cells can be useful for propagation of HDDPCs by eliminating contaminating bacteria and by promoting cellular outgrowth.
{"title":"STO Feeder Cells Are Useful for Propagation of Primarily Cultured Human Deciduous Dental Pulp Cells by Eliminating Contaminating Bacteria and Promoting Cellular Outgrowth.","authors":"Tomoya Murakami, I. Saitoh, E. Inada, Mie Kurosawa, Y. Iwase, H. Noguchi, Y. Terao, Y. Yamasaki, H. Hayasaki, Masahiro Sato","doi":"10.3727/215517913X674234","DOIUrl":"https://doi.org/10.3727/215517913X674234","url":null,"abstract":"STO feeder cells, a line established from mouse SIM embryonic fibroblasts, have been frequently used for establishing embryonic stem cells and maintaining them in an undifferentiated state. There are some reports demonstrating that fibroblastic cells have the ability to phagocytose Gram-positive bacterium (e.g., streptococci and staphylococci). In this study, we examined the possibility that STO cells could phagocytose Streptococcus mutans (a bacteria causing tooth decay), which always contaminates cultures of primarily isolated human deciduous dental pulp cells (HDDPCs). Simple cultivation of the primary HDDPCs in the absence of STO cells allowed S. mutans to massively propagate in the medium, thus leading to an opaque medium. In contrast, there was no bacterial contamination in the cultures containing mitomycin C (MMC)-inactivated STO cells. Furthermore, STO cells indicated bacterial phagocytic activity under fluorescent microscopy with the dye pHrodo. Besides removal of contaminating bacteria, STO feeder cells allowed the HDDPCs to spread out. These data suggest that MMC-treated STO cells can be useful for propagation of HDDPCs by eliminating contaminating bacteria and by promoting cellular outgrowth.","PeriodicalId":9780,"journal":{"name":"Cell medicine","volume":"6 1-2 1","pages":"75-81"},"PeriodicalIF":0.0,"publicationDate":"2013-12-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3727/215517913X674234","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69756244","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 : 2013-12-30DOI: 10.3727/215517913X674243
Shingo Yamashita, K. Ohashi, R. Utoh, T. Kin, A. M. J. Shapiro, Masakazu Yamamoto, M. Gotoh, T. Okano
In new generation medical therapies for type 1 diabetes mellitus (DM), cell-based approaches using pancreatic islets have attracted significant attention worldwide. In particular, dispersed islet cells obtained from isolated pancreatic islets have been a valuable source in the cell biology and tissue engineering fields. Our experimental approach to the development of new islet-based DM therapies consisted of creating a monolithic islet cell sheet format using dispersed islet cells. In this experiment, we explored the potential of internationally transporting human islets from Alberta, Canada to Tokyo, Japan and obtaining viable dispersed islet cells. A total of 34 batches of isolated and purified human islets were transported using a commercial air courier service. Prior to shipping, the human islets had been in culture for 0-108 h at the University of Alberta. The transportation period from Alberta to Tokyo was 2-5 days. The transported human islet cells were enzymatically dispersed as single cells in Tokyo. The number of single islet cells decreased as the number of transportation days increased. In contrast, cell viability was maintained regardless of the number of transportation days. The preshipment culture time had no effect on the number or viability of single cells dispersed in Tokyo. When dispersed single islet cells were plated on laminin-5-coated temperature-responsive polymer-grafted culture dishes, the cells showed favorable attachment followed by extension as a monolithic format. The present study demonstrated that long-distance transported human islets are a viable cell source for experiments utilizing dispersed human islet cells.
{"title":"Quality of Air-Transported Human Islets for Single Islet Cell Preparations.","authors":"Shingo Yamashita, K. Ohashi, R. Utoh, T. Kin, A. M. J. Shapiro, Masakazu Yamamoto, M. Gotoh, T. Okano","doi":"10.3727/215517913X674243","DOIUrl":"https://doi.org/10.3727/215517913X674243","url":null,"abstract":"In new generation medical therapies for type 1 diabetes mellitus (DM), cell-based approaches using pancreatic islets have attracted significant attention worldwide. In particular, dispersed islet cells obtained from isolated pancreatic islets have been a valuable source in the cell biology and tissue engineering fields. Our experimental approach to the development of new islet-based DM therapies consisted of creating a monolithic islet cell sheet format using dispersed islet cells. In this experiment, we explored the potential of internationally transporting human islets from Alberta, Canada to Tokyo, Japan and obtaining viable dispersed islet cells. A total of 34 batches of isolated and purified human islets were transported using a commercial air courier service. Prior to shipping, the human islets had been in culture for 0-108 h at the University of Alberta. The transportation period from Alberta to Tokyo was 2-5 days. The transported human islet cells were enzymatically dispersed as single cells in Tokyo. The number of single islet cells decreased as the number of transportation days increased. In contrast, cell viability was maintained regardless of the number of transportation days. The preshipment culture time had no effect on the number or viability of single cells dispersed in Tokyo. When dispersed single islet cells were plated on laminin-5-coated temperature-responsive polymer-grafted culture dishes, the cells showed favorable attachment followed by extension as a monolithic format. The present study demonstrated that long-distance transported human islets are a viable cell source for experiments utilizing dispersed human islet cells.","PeriodicalId":9780,"journal":{"name":"Cell medicine","volume":"6 1-2 1","pages":"33-8"},"PeriodicalIF":0.0,"publicationDate":"2013-12-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3727/215517913X674243","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69756253","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 : 2013-11-10DOI: 10.3727/215517913X666512
H. Kataoka, H. Noguchi
Endoplasmic reticulum (ER) stress affects the pathogenesis of diabetes. ER stress plays important roles, both in type 1 and type 2 diabetes, because pancreatic β-cells possess highly developed ER for insulin secretion. This review summarizes the relationship between ER stress and the pathogenesis of type 1 and type 2 diabetes. In addition, the association between islet transplantation and ER stress is discussed.
{"title":"ER Stress and β-Cell Pathogenesis of Type 1 and Type 2 Diabetes and Islet Transplantation.","authors":"H. Kataoka, H. Noguchi","doi":"10.3727/215517913X666512","DOIUrl":"https://doi.org/10.3727/215517913X666512","url":null,"abstract":"Endoplasmic reticulum (ER) stress affects the pathogenesis of diabetes. ER stress plays important roles, both in type 1 and type 2 diabetes, because pancreatic β-cells possess highly developed ER for insulin secretion. This review summarizes the relationship between ER stress and the pathogenesis of type 1 and type 2 diabetes. In addition, the association between islet transplantation and ER stress is discussed.","PeriodicalId":9780,"journal":{"name":"Cell medicine","volume":"5 2-3 1","pages":"53-7"},"PeriodicalIF":0.0,"publicationDate":"2013-11-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3727/215517913X666512","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69756191","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 : 2013-11-10DOI: 10.3727/215517913X666486
M. Dozen, S. Enosawa, Yuki Tada, K. Hirasawa
The pathogenesis of ischemia/reperfusion (I/R) injury in surgical trauma, organ transplantations, and brain and myocardial infarctions is attributable to excessive oxidative stress caused by reactive oxygen species and their metabolites. We prepared a physiological saline solution treated with simple exposure to a microampere current with electron discharge onto the surface; this treatment increased reduction potential and caused proton reduction. We examined the reductive potency of the electron-treated solution (ETS) on hepatocellular I/R injury in a rat model. When the ETS was administered in four doses at 0, 3, 10, and 20 min after reperfusion, severe hepatocellular injury was suppressed, as revealed by a decrease in serum alanine aminotransferase levels and histopathological improvement of liver damage. Since a preparation of hydrogen gas-dissolved saline solution (HDS) was shown to be capable of suppressing I/R injury, the possible involvement of dissolved hydrogen gas in the effectiveness of ETS was examined. When HDS was treated by degasification, the aforementioned effectiveness was decreased. In contrast, the same treatment did not alter the effectiveness of ETS. These results suggest that the antioxidative efficacy of ETS is not attributable to dissolved hydrogen gas but presumably to the molecule(s) produced from the stepwise reduction of protons in the solution.
{"title":"Inhibition of Hepatic Ischemic Reperfusion Injury Using Saline Exposed to Electron Discharge in a Rat Model.","authors":"M. Dozen, S. Enosawa, Yuki Tada, K. Hirasawa","doi":"10.3727/215517913X666486","DOIUrl":"https://doi.org/10.3727/215517913X666486","url":null,"abstract":"The pathogenesis of ischemia/reperfusion (I/R) injury in surgical trauma, organ transplantations, and brain and myocardial infarctions is attributable to excessive oxidative stress caused by reactive oxygen species and their metabolites. We prepared a physiological saline solution treated with simple exposure to a microampere current with electron discharge onto the surface; this treatment increased reduction potential and caused proton reduction. We examined the reductive potency of the electron-treated solution (ETS) on hepatocellular I/R injury in a rat model. When the ETS was administered in four doses at 0, 3, 10, and 20 min after reperfusion, severe hepatocellular injury was suppressed, as revealed by a decrease in serum alanine aminotransferase levels and histopathological improvement of liver damage. Since a preparation of hydrogen gas-dissolved saline solution (HDS) was shown to be capable of suppressing I/R injury, the possible involvement of dissolved hydrogen gas in the effectiveness of ETS was examined. When HDS was treated by degasification, the aforementioned effectiveness was decreased. In contrast, the same treatment did not alter the effectiveness of ETS. These results suggest that the antioxidative efficacy of ETS is not attributable to dissolved hydrogen gas but presumably to the molecule(s) produced from the stepwise reduction of protons in the solution.","PeriodicalId":9780,"journal":{"name":"Cell medicine","volume":"5 2-3 1","pages":"83-7"},"PeriodicalIF":0.0,"publicationDate":"2013-11-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3727/215517913X666486","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69756100","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 : 2013-10-29DOI: 10.3727/215517913X674261
Yoshiyuki Miyazaki, H. Yukawa, Hiroyasu Nishi, Y. Okamoto, N. Kaji, T. Torimoto, Y. Baba
Quantum dots (QDs) have received much attention for biomolecule and cell imaging applications because of their superior optical properties such as high quantum efficiency, size-tunable emission, and resistance to photobleaching process. However, QDs that are commercially available contain cadmium (Cd), a highly toxic element. Thus, the development of Cd-free and less toxic QDs is strongly desired. In this study, we developed Cd-free QDs (ZnS-coated ZnS-AgInS2 solid solution nanoparticles with a sulfo group: ZnS-ZAIS-SO3H) and investigated the ability of this material to label stem cells. ZnS-ZAIS-SO3H could be transduced into mouse adipose tissue-derived stem cells (mASCs) using octaarginine peptides (R8), known as cell-penetrating peptides. The optimal ratio of ZnS-ZAIS-SO3H:R8 was found to be 1:100 for labeling mASCs. More than 80% of mASCs labeled with 500 nM ZnS-ZAIS-SO3H were found to be alive, and the proliferation rates of labeled mASCs were maintained at the same rate as that of nonlabeled mASCs. In addition, no abnormalities in the morphology of mASCs labeled with ZnS-ZAIS-SO3H could be observed. These data suggest that ZnS-ZAIS-SO3H may be effective for the labeling of mASCs.
{"title":"Adipose Tissue-Derived Stem Cell Imaging Using Cadmium-Free Quantum Dots.","authors":"Yoshiyuki Miyazaki, H. Yukawa, Hiroyasu Nishi, Y. Okamoto, N. Kaji, T. Torimoto, Y. Baba","doi":"10.3727/215517913X674261","DOIUrl":"https://doi.org/10.3727/215517913X674261","url":null,"abstract":"Quantum dots (QDs) have received much attention for biomolecule and cell imaging applications because of their superior optical properties such as high quantum efficiency, size-tunable emission, and resistance to photobleaching process. However, QDs that are commercially available contain cadmium (Cd), a highly toxic element. Thus, the development of Cd-free and less toxic QDs is strongly desired. In this study, we developed Cd-free QDs (ZnS-coated ZnS-AgInS2 solid solution nanoparticles with a sulfo group: ZnS-ZAIS-SO3H) and investigated the ability of this material to label stem cells. ZnS-ZAIS-SO3H could be transduced into mouse adipose tissue-derived stem cells (mASCs) using octaarginine peptides (R8), known as cell-penetrating peptides. The optimal ratio of ZnS-ZAIS-SO3H:R8 was found to be 1:100 for labeling mASCs. More than 80% of mASCs labeled with 500 nM ZnS-ZAIS-SO3H were found to be alive, and the proliferation rates of labeled mASCs were maintained at the same rate as that of nonlabeled mASCs. In addition, no abnormalities in the morphology of mASCs labeled with ZnS-ZAIS-SO3H could be observed. These data suggest that ZnS-ZAIS-SO3H may be effective for the labeling of mASCs.","PeriodicalId":9780,"journal":{"name":"Cell medicine","volume":"6 1-2 1","pages":"91-7"},"PeriodicalIF":0.0,"publicationDate":"2013-10-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3727/215517913X674261","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69756314","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 : 2013-10-23DOI: 10.3727/215517913X674225
S. Enosawa, M. Dozen, Yuki Tada, K. Hirasawa
Chronic oxidative stress plays a key role in the progression of nonalcoholic steatohepatitis (NASH). We examined the efficacy of antioxidative electron treatment on type 2 diabetes-induced NASH in a rat model. We established NASH model rats, induced by neonatal administration of streptozotocin and a high-fat diet, which exhibited pathologically high values of alanine aminotransferase (ALT), glucose, and malondialdehyde (MDA). The rats were exposed to electron discharge at very low energy for 4 weeks; this dose results in the reduction of Fe(3+) and glutathione disulfide in vitro. Serum ALT values were increased from baseline (8 weeks) to 125.0 ± 13 U/L at 10 weeks in the control group. In contrast, the values in the treated group did not show any increase at 10 weeks [87 ± 10 U/L (p = 0.0391)]. Hepatic MDA levels were also significantly decreased at 12 weeks (p < 0.05), but 8-hydroxy-2'-deoxyguanosine values were not statistically significant (p = 0.076). A gradual but steadily decreasing trend from initially high glucose levels was observed, though the values were not significant in 12-week-old animals (p = 0.074). However, the serum values of MDA, ALT, and glucose were well correlated. The progression of fibrosis as measured by increased serum levels of hyaluronic acid and histological examinations were not affected by the treatment in this model. Antioxidative electron treatment at very low energy attenuated the pathogenically elevated liver inflammation and oxidative stress, together with presumably impaired glucose metabolism in NASH rat model.
{"title":"Electron Therapy Attenuated Elevated Alanine Aminotransferase and Oxidative Stress Values in Type 2 Diabetes-Induced Nonalcoholic Steatohepatitis of Rats.","authors":"S. Enosawa, M. Dozen, Yuki Tada, K. Hirasawa","doi":"10.3727/215517913X674225","DOIUrl":"https://doi.org/10.3727/215517913X674225","url":null,"abstract":"Chronic oxidative stress plays a key role in the progression of nonalcoholic steatohepatitis (NASH). We examined the efficacy of antioxidative electron treatment on type 2 diabetes-induced NASH in a rat model. We established NASH model rats, induced by neonatal administration of streptozotocin and a high-fat diet, which exhibited pathologically high values of alanine aminotransferase (ALT), glucose, and malondialdehyde (MDA). The rats were exposed to electron discharge at very low energy for 4 weeks; this dose results in the reduction of Fe(3+) and glutathione disulfide in vitro. Serum ALT values were increased from baseline (8 weeks) to 125.0 ± 13 U/L at 10 weeks in the control group. In contrast, the values in the treated group did not show any increase at 10 weeks [87 ± 10 U/L (p = 0.0391)]. Hepatic MDA levels were also significantly decreased at 12 weeks (p < 0.05), but 8-hydroxy-2'-deoxyguanosine values were not statistically significant (p = 0.076). A gradual but steadily decreasing trend from initially high glucose levels was observed, though the values were not significant in 12-week-old animals (p = 0.074). However, the serum values of MDA, ALT, and glucose were well correlated. The progression of fibrosis as measured by increased serum levels of hyaluronic acid and histological examinations were not affected by the treatment in this model. Antioxidative electron treatment at very low energy attenuated the pathogenically elevated liver inflammation and oxidative stress, together with presumably impaired glucose metabolism in NASH rat model.","PeriodicalId":9780,"journal":{"name":"Cell medicine","volume":"39 1","pages":"63-73"},"PeriodicalIF":0.0,"publicationDate":"2013-10-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3727/215517913X674225","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69756684","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 : 2013-10-23DOI: 10.3727/215517913X674216
K. Sugiyama, H. Sasahara, M. Tsukaguchi, K. Isogai, A. Toyama, H. Satoh, K. Saitoh, Y. Nakagawa, Kota Takahashi, Sachiko Tanaka, K. Onda, T. Hirano
The lymphocyte immunosuppressant sensitivity test (LIST) with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay procedure has been used to predict the pharmacological efficacy of immunosuppressive agents to prevent acute rejection episodes for renal transplant recipients. In this study, mycophenolic acid (MPA) pharmacological efficacies were evaluated by LIST at both prior to and just after renal transplantation. We compared the efficacies to the clinical outcome of these recipients. MPA's pharmacological efficacy was evaluated by LIST not only before the operation but also at 2, 4, and 6 weeks after transplantation in 16 renal transplant recipients. These recipients were divided into high- and low-sensitivity groups according to peripheral blood mononuclear cell (PBMC) sensitivity to MPA in vitro. The MPA sensitivities were compared to cytomegalovirus (CMV) infection and acute rejection episodes in these recipients under MPA immunosuppressive therapy. The rate of CMV infection episodes in the low-MPA pharmacological efficacy group categorized at 2 weeks after renal transplantation was 5/6 (83.3%), which was significantly higher than the rate of 1/10 (10.0%) (p < 0.01) in the high-MPA sensitivity group. However, the MPA pharmacological efficacy evaluated both before and after transplantation had no relationship with the incidence of rejection episodes. These findings suggest that the MPA pharmacological efficacy evaluated by LIST at 2 weeks after operation is a useful biomarker for predicting the following occurrence of CMV infection episodes in renal transplant recipients.
{"title":"Peripheral Lymphocyte Response to Mycophenolic Acid In Vitro and Incidence of Cytomegalovirus Infection in Renal Transplantation.","authors":"K. Sugiyama, H. Sasahara, M. Tsukaguchi, K. Isogai, A. Toyama, H. Satoh, K. Saitoh, Y. Nakagawa, Kota Takahashi, Sachiko Tanaka, K. Onda, T. Hirano","doi":"10.3727/215517913X674216","DOIUrl":"https://doi.org/10.3727/215517913X674216","url":null,"abstract":"The lymphocyte immunosuppressant sensitivity test (LIST) with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay procedure has been used to predict the pharmacological efficacy of immunosuppressive agents to prevent acute rejection episodes for renal transplant recipients. In this study, mycophenolic acid (MPA) pharmacological efficacies were evaluated by LIST at both prior to and just after renal transplantation. We compared the efficacies to the clinical outcome of these recipients. MPA's pharmacological efficacy was evaluated by LIST not only before the operation but also at 2, 4, and 6 weeks after transplantation in 16 renal transplant recipients. These recipients were divided into high- and low-sensitivity groups according to peripheral blood mononuclear cell (PBMC) sensitivity to MPA in vitro. The MPA sensitivities were compared to cytomegalovirus (CMV) infection and acute rejection episodes in these recipients under MPA immunosuppressive therapy. The rate of CMV infection episodes in the low-MPA pharmacological efficacy group categorized at 2 weeks after renal transplantation was 5/6 (83.3%), which was significantly higher than the rate of 1/10 (10.0%) (p < 0.01) in the high-MPA sensitivity group. However, the MPA pharmacological efficacy evaluated both before and after transplantation had no relationship with the incidence of rejection episodes. These findings suggest that the MPA pharmacological efficacy evaluated by LIST at 2 weeks after operation is a useful biomarker for predicting the following occurrence of CMV infection episodes in renal transplant recipients.","PeriodicalId":9780,"journal":{"name":"Cell medicine","volume":"6 1-2 1","pages":"47-55"},"PeriodicalIF":0.0,"publicationDate":"2013-10-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3727/215517913X674216","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69756675","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}
京公网安备 11010802042870号
京ICP备2023020795号-1
扫码关注我们
求助内容：
应助结果提醒方式：