Alicja Szubarga, M. Kamińska, Wiktoria Kotlarz, Stefan Malewski, Wiktoria Zawada, Matylda Kuczma, M. Ješeta, P. Antosik
Abstract Stem cells are an important subject of research, and are increasingly used in the treatment of various diseases. Due to the development of advanced in vitro techniques, they have become an integral part of modern medicine. The sources of human stem cells are primarily bone marrow and adipose tissue, although non – embryonic stem cells are also scattered throughout the body. Notably, recent research has focused on stem cells found in the oral cavity, both in the dental pulp and oral mucosa. Furthermore, isolation of stem cells from umbilical cord blood is also becoming increasingly popular, while wharton’s jelly and amniotic fluid also seem to be an interesting source of stem cells. The safety and efficacy of stem cells use can be established by animal studies, which are a key element of preclinical research. Mouse, rat and pig models allow for testing of stem cell therapies. Recent studies primarily use mesenchymal stem cells such as mouse – adipose derived mesenchymal stem cells and mouse and rat hematopoietic stem cells. Great hope for future therapies is the use of bioengineering to program cells into induced stem cells, which have the biggest ability for differentiation and transdifferentiation, which carries no risk of teratogenesis. Stem cells are used in many areas of medicine, especially in regenerative medicine, with a growing interest in orthopedics and in the treatment of heart failure. Mesenchymal stem cells are the most used stem cell type, which despite their limited ability to differentiate, give great therapeutic results, mainly due to their immunomodulating effect. Recent studies have even shown that the use of mesenchymal stem cells may be useful in the treatment of COVID-19. Moreover, Research on the use of mesenchymal stem cells in the treatment of Crohn’s disease, acute-graft-versus-host disease and type I diabetes are also promising. The aim of the current review is to present and systematize current knowledge about stem cells, their use and related in vitro research. Running title: Research and use of human stem cells
{"title":"Human stem cells – sources, sourcing and in vitro methods","authors":"Alicja Szubarga, M. Kamińska, Wiktoria Kotlarz, Stefan Malewski, Wiktoria Zawada, Matylda Kuczma, M. Ješeta, P. Antosik","doi":"10.2478/acb-2021-0011","DOIUrl":"https://doi.org/10.2478/acb-2021-0011","url":null,"abstract":"Abstract Stem cells are an important subject of research, and are increasingly used in the treatment of various diseases. Due to the development of advanced in vitro techniques, they have become an integral part of modern medicine. The sources of human stem cells are primarily bone marrow and adipose tissue, although non – embryonic stem cells are also scattered throughout the body. Notably, recent research has focused on stem cells found in the oral cavity, both in the dental pulp and oral mucosa. Furthermore, isolation of stem cells from umbilical cord blood is also becoming increasingly popular, while wharton’s jelly and amniotic fluid also seem to be an interesting source of stem cells. The safety and efficacy of stem cells use can be established by animal studies, which are a key element of preclinical research. Mouse, rat and pig models allow for testing of stem cell therapies. Recent studies primarily use mesenchymal stem cells such as mouse – adipose derived mesenchymal stem cells and mouse and rat hematopoietic stem cells. Great hope for future therapies is the use of bioengineering to program cells into induced stem cells, which have the biggest ability for differentiation and transdifferentiation, which carries no risk of teratogenesis. Stem cells are used in many areas of medicine, especially in regenerative medicine, with a growing interest in orthopedics and in the treatment of heart failure. Mesenchymal stem cells are the most used stem cell type, which despite their limited ability to differentiate, give great therapeutic results, mainly due to their immunomodulating effect. Recent studies have even shown that the use of mesenchymal stem cells may be useful in the treatment of COVID-19. Moreover, Research on the use of mesenchymal stem cells in the treatment of Crohn’s disease, acute-graft-versus-host disease and type I diabetes are also promising. The aim of the current review is to present and systematize current knowledge about stem cells, their use and related in vitro research. Running title: Research and use of human stem cells","PeriodicalId":18329,"journal":{"name":"Medical Journal of Cell Biology","volume":"9 1","pages":"73 - 85"},"PeriodicalIF":0.0,"publicationDate":"2021-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49492056","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}
Rafał Sibiak, Rut Bryl, K. Stefańska, B. Chermuła, W. Pieńkowski, M. Ješeta, L. Pawelczyk, P. Mozdziak, R. Spaczyński, B. Kempisty
Abstract Human cumulus cells (CCs) play a key role in the regulation of ovarian follicle maturation and oocyte fertilization. They influence the oocyte development by transferring the various molecules via the specific gap junction proteins, also known as the connexins, which provide a direct transmembrane connection between the oocyte and CCs. The human CCs were obtained in the patients diagnosed with infertility, who underwent the procedure of the controlled ovarian stimulation, and the following in vitro fertilization to elucidate the possible involvement of the CCs in the regulation of the fertilization and oocyte aging. Collected samples were long-term cultured and harvested after 7, 15, and 30 days of cultivation. Afterward, we assessed the relative expression of the following apoptosis regulatory genes - BAX, CASP9, and TP53 - using the RT-qPCR method. We noted a decrease in the expression of all above-mentioned genes in the samples harvested after 15 and 30 days, in reference to 7 days in vitro cultured CCs. In summary, our results provide precious insight into the dynamics of changes and confirm the continuous expression of the proapoptotic genes – BAX, CASP9, and TP53 in the long-term cultured CCs. Running title: Apoptotic gene expression in the human cumulus cells
{"title":"Expression of the apoptosis regulatory gene family in the long-term in vitro cultured human cumulus cells","authors":"Rafał Sibiak, Rut Bryl, K. Stefańska, B. Chermuła, W. Pieńkowski, M. Ješeta, L. Pawelczyk, P. Mozdziak, R. Spaczyński, B. Kempisty","doi":"10.2478/acb-2021-0002","DOIUrl":"https://doi.org/10.2478/acb-2021-0002","url":null,"abstract":"Abstract Human cumulus cells (CCs) play a key role in the regulation of ovarian follicle maturation and oocyte fertilization. They influence the oocyte development by transferring the various molecules via the specific gap junction proteins, also known as the connexins, which provide a direct transmembrane connection between the oocyte and CCs. The human CCs were obtained in the patients diagnosed with infertility, who underwent the procedure of the controlled ovarian stimulation, and the following in vitro fertilization to elucidate the possible involvement of the CCs in the regulation of the fertilization and oocyte aging. Collected samples were long-term cultured and harvested after 7, 15, and 30 days of cultivation. Afterward, we assessed the relative expression of the following apoptosis regulatory genes - BAX, CASP9, and TP53 - using the RT-qPCR method. We noted a decrease in the expression of all above-mentioned genes in the samples harvested after 15 and 30 days, in reference to 7 days in vitro cultured CCs. In summary, our results provide precious insight into the dynamics of changes and confirm the continuous expression of the proapoptotic genes – BAX, CASP9, and TP53 in the long-term cultured CCs. Running title: Apoptotic gene expression in the human cumulus cells","PeriodicalId":18329,"journal":{"name":"Medical Journal of Cell Biology","volume":"9 1","pages":"8 - 13"},"PeriodicalIF":0.0,"publicationDate":"2021-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45228559","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}
Paulina Pietras, Marta Leśniczak, Mateusz Sowiński, Witold Szaflarski
Abstract Stress granules (SGs) are cytoplasmic structures found in eukaryotic cells, from yeast to human cells. They are made up of proteins, RNA and small ribosome subunits (40S). They arise as a result of the rapid shutdown of active protein biosynthesis in the cell, which is the result of the appearance of a stress factor. The mechanism of regulation of protein biosynthesis in response to stress takes place at two control nodes: (1) phosphorylation of the α subunit of the eIF2 factor as a result of the action of stress-recognizing kinases or by modulation of the mTOR pathway activity, which regulates the initiation of protein biosynthesis by the formation of a complex within the so-called cap structure. The protein arrest causes aggregation of the translation process components and other cell components (other proteins or mRNA molecules) into SGs. A lot of data indicates the active participation of SGs in metabolic processes, their control role over pro- and anti-apoptotic processes as well as in the development of cancer, neurodegenerative diseases and their defensive role in viral infections. Running title: Stress granules in the cell
{"title":"Molecular structure of stress granules and their role in the eukaryotic cell","authors":"Paulina Pietras, Marta Leśniczak, Mateusz Sowiński, Witold Szaflarski","doi":"10.2478/acb-2021-0006","DOIUrl":"https://doi.org/10.2478/acb-2021-0006","url":null,"abstract":"Abstract Stress granules (SGs) are cytoplasmic structures found in eukaryotic cells, from yeast to human cells. They are made up of proteins, RNA and small ribosome subunits (40S). They arise as a result of the rapid shutdown of active protein biosynthesis in the cell, which is the result of the appearance of a stress factor. The mechanism of regulation of protein biosynthesis in response to stress takes place at two control nodes: (1) phosphorylation of the α subunit of the eIF2 factor as a result of the action of stress-recognizing kinases or by modulation of the mTOR pathway activity, which regulates the initiation of protein biosynthesis by the formation of a complex within the so-called cap structure. The protein arrest causes aggregation of the translation process components and other cell components (other proteins or mRNA molecules) into SGs. A lot of data indicates the active participation of SGs in metabolic processes, their control role over pro- and anti-apoptotic processes as well as in the development of cancer, neurodegenerative diseases and their defensive role in viral infections. Running title: Stress granules in the cell","PeriodicalId":18329,"journal":{"name":"Medical Journal of Cell Biology","volume":"9 1","pages":"33 - 41"},"PeriodicalIF":0.0,"publicationDate":"2021-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41354429","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}
A. G. Narenji, J. Petitte, M. Kulus, K. Stefańska, Joanna Perek, Jakub Kulus, M. Wieczorkiewicz, M. Jankowski, A. Bryja, Rut Bryl, P. Antosik, D. Bukowska, Jędrzej M. Jaśkowki, Kornel Ratajczak, P. Mozdziak
Abstract Telomerase activity is highly correlated to the proliferation capacity and immortality of cells. To evaluate the possibility of continuous culture, myoblasts were isolated from the Pectoralis thoracicus muscle of newborn turkeys and maintained in 2D (adherence based) and suspension cultures. Furthermore, adherent myoblasts were differentiated into myotubes. Telomerase activity was evaluated in all types of obtained cultures. The expression of telomerase related genes, including TERT1, TERT2, dyskerin, as well as myogenesis related genes, including myogenin, MyoD, MRF1 and MRF5 were measured. Telomerase bands were detected in both adherent and suspended cells, but they were not detected in samples from rat muscle. Myotube differentiation caused a significant reduction in the expression of TERT1, TERT2 and Dyskerin, while MyoD, Myogenin and MRF4 were upregulated in myotubes vs. myoblasts. Long-term culture of suspended myoblasts caused a significant increase in TERT1 levels, with no significant change in expression of myogenesis related genes. Overall, the results show that myoblasts are able to grow in suspension without losing their myogenic properties. Furthermore, upregulation of TERT1 indicates continued proliferation of myoblasts and generation of enough daughter cells necessary for in vitro meat production. Running title: Telomerase activity and myogenic properties of cultured Turkey satellite cells
{"title":"Telomerase Activity and Myogenesis Ability as an Indicator of Cultured Turkey Satellite Cell Ability for In Vitro Meat Production","authors":"A. G. Narenji, J. Petitte, M. Kulus, K. Stefańska, Joanna Perek, Jakub Kulus, M. Wieczorkiewicz, M. Jankowski, A. Bryja, Rut Bryl, P. Antosik, D. Bukowska, Jędrzej M. Jaśkowki, Kornel Ratajczak, P. Mozdziak","doi":"10.2478/acb-2021-0004","DOIUrl":"https://doi.org/10.2478/acb-2021-0004","url":null,"abstract":"Abstract Telomerase activity is highly correlated to the proliferation capacity and immortality of cells. To evaluate the possibility of continuous culture, myoblasts were isolated from the Pectoralis thoracicus muscle of newborn turkeys and maintained in 2D (adherence based) and suspension cultures. Furthermore, adherent myoblasts were differentiated into myotubes. Telomerase activity was evaluated in all types of obtained cultures. The expression of telomerase related genes, including TERT1, TERT2, dyskerin, as well as myogenesis related genes, including myogenin, MyoD, MRF1 and MRF5 were measured. Telomerase bands were detected in both adherent and suspended cells, but they were not detected in samples from rat muscle. Myotube differentiation caused a significant reduction in the expression of TERT1, TERT2 and Dyskerin, while MyoD, Myogenin and MRF4 were upregulated in myotubes vs. myoblasts. Long-term culture of suspended myoblasts caused a significant increase in TERT1 levels, with no significant change in expression of myogenesis related genes. Overall, the results show that myoblasts are able to grow in suspension without losing their myogenic properties. Furthermore, upregulation of TERT1 indicates continued proliferation of myoblasts and generation of enough daughter cells necessary for in vitro meat production. Running title: Telomerase activity and myogenic properties of cultured Turkey satellite cells","PeriodicalId":18329,"journal":{"name":"Medical Journal of Cell Biology","volume":"9 1","pages":"19 - 26"},"PeriodicalIF":0.0,"publicationDate":"2021-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46571793","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}
Zeinab Shafiei Seifabadi, Fatemeh Rezaei-Tazangi, Nastaran Azarbarz, D. Nejad, J. Mohammadiasl, Hanieh Darabi, Seyed Pezhmanlarki-Tork
Abstract Cell encapsulation utilizing biodegradable material has promising outcomes for tissue engineering. From a long time ago, alginate has been generally utilized for drug delivery, cell transplantation and as a scaffold in biomedical applications. The aim of this study was the comparison of cell viability in the presence of two polymerizing ions: Ba2+ and Ca2+ to improvement the quality of alginate scaffold. For this purpose, WJMSCs after three passage were encapsulated in alginate scaffold in the presence of Ba2+ and ca2+. Cell viability was evaluated by WST-8 assay kit after 24, 48 and 72 hours. The results showed that encapsulated cells in the presence of Ca2+ had more viability than Ba2+. It was also found that using the WST-8 assay kit is a convenient and fast method for evaluation the viability of cells. It can be claimed that Calcl2 polymerizing solution provides more favorable conditions for cell viability compared to Bacl2 solution. Running title: Assessing the viability of stem cells by WST-8 assay kit
{"title":"Assessment of viability of wharton's jelly mesenchymal stem cells encapsulated in alginate scaffold by WST-8 assay kit","authors":"Zeinab Shafiei Seifabadi, Fatemeh Rezaei-Tazangi, Nastaran Azarbarz, D. Nejad, J. Mohammadiasl, Hanieh Darabi, Seyed Pezhmanlarki-Tork","doi":"10.2478/acb-2021-0007","DOIUrl":"https://doi.org/10.2478/acb-2021-0007","url":null,"abstract":"Abstract Cell encapsulation utilizing biodegradable material has promising outcomes for tissue engineering. From a long time ago, alginate has been generally utilized for drug delivery, cell transplantation and as a scaffold in biomedical applications. The aim of this study was the comparison of cell viability in the presence of two polymerizing ions: Ba2+ and Ca2+ to improvement the quality of alginate scaffold. For this purpose, WJMSCs after three passage were encapsulated in alginate scaffold in the presence of Ba2+ and ca2+. Cell viability was evaluated by WST-8 assay kit after 24, 48 and 72 hours. The results showed that encapsulated cells in the presence of Ca2+ had more viability than Ba2+. It was also found that using the WST-8 assay kit is a convenient and fast method for evaluation the viability of cells. It can be claimed that Calcl2 polymerizing solution provides more favorable conditions for cell viability compared to Bacl2 solution. Running title: Assessing the viability of stem cells by WST-8 assay kit","PeriodicalId":18329,"journal":{"name":"Medical Journal of Cell Biology","volume":"9 1","pages":"42 - 47"},"PeriodicalIF":0.0,"publicationDate":"2021-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42374064","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}
Rut Bryl, K. Stefańska, B. Chermuła, B. Stelmach, W. Pieńkowski, Jakub Kulus, Joanna Perek, M. Wieczorkiewicz, G. Wąsiatycz, Kornel Ratajczak, L. Pawelczyk, P. Mozdziak, M. Ješeta, R. Spaczyński, D. Bukowska
Abstract Granulosa cells play an important role in follicle development, maturation, and atresia. They are a cellular source of the two most important ovarian steroids, namely, estradiol and progesterone and are also crucial for bidirectional communication with the oocyte, thus being involved in the regulation of its growth, development and function. Growing body of evidence suggests that granulosa cells cultured in vitro display stemness and transdifferentiation potential. Together with the fact that they can be easily collected during IVF procedures, these properties of GCs may be of particular interest for both regenerative medicine and transplantology. Establishment of in vitro cell culture and its thorough characterization, including molecular, is crucial for future potential utilization of human granulosa cells in design of engineered tissue grafts or cell-based therapies, in particular targeted at female infertility. Nevertheless, the transcriptomic alterations which may occur during in vitro culture of granulosa cells are still largely uncharacterized. The aim of this study was to examine expression changes of three genes encoding histone demethylases which serve as transcription coactivators in short term in vitro cell culture of human granulosa cells. The study groups consisted of 14 patients, aged 18–40 years undergoing in vitro fertilization (IVF). Expression level assessment was performed after 24 h, 48 h, 72 h, 96 h, 120 h, 144 h and 168 h of in vitro primary cell culture utilizing RT-qPCR technique. Upregulation of PHF2 expression in all time points of the culture was observed, whereas the tendency of JHDM1D and PHF8 was mainly to decrease in expression level. Further study on a larger population would be required in order to confirm the presented tendencies. Running title: Expression pattern of selected histone demethylases in human granulosa cells
{"title":"Histone demethylases JHDM1D, PHF2 and PHF8 expression pattern in granulosa cells obtained from patients undergoing IVF procedure during short-term IVC","authors":"Rut Bryl, K. Stefańska, B. Chermuła, B. Stelmach, W. Pieńkowski, Jakub Kulus, Joanna Perek, M. Wieczorkiewicz, G. Wąsiatycz, Kornel Ratajczak, L. Pawelczyk, P. Mozdziak, M. Ješeta, R. Spaczyński, D. Bukowska","doi":"10.2478/acb-2021-0001","DOIUrl":"https://doi.org/10.2478/acb-2021-0001","url":null,"abstract":"Abstract Granulosa cells play an important role in follicle development, maturation, and atresia. They are a cellular source of the two most important ovarian steroids, namely, estradiol and progesterone and are also crucial for bidirectional communication with the oocyte, thus being involved in the regulation of its growth, development and function. Growing body of evidence suggests that granulosa cells cultured in vitro display stemness and transdifferentiation potential. Together with the fact that they can be easily collected during IVF procedures, these properties of GCs may be of particular interest for both regenerative medicine and transplantology. Establishment of in vitro cell culture and its thorough characterization, including molecular, is crucial for future potential utilization of human granulosa cells in design of engineered tissue grafts or cell-based therapies, in particular targeted at female infertility. Nevertheless, the transcriptomic alterations which may occur during in vitro culture of granulosa cells are still largely uncharacterized. The aim of this study was to examine expression changes of three genes encoding histone demethylases which serve as transcription coactivators in short term in vitro cell culture of human granulosa cells. The study groups consisted of 14 patients, aged 18–40 years undergoing in vitro fertilization (IVF). Expression level assessment was performed after 24 h, 48 h, 72 h, 96 h, 120 h, 144 h and 168 h of in vitro primary cell culture utilizing RT-qPCR technique. Upregulation of PHF2 expression in all time points of the culture was observed, whereas the tendency of JHDM1D and PHF8 was mainly to decrease in expression level. Further study on a larger population would be required in order to confirm the presented tendencies. Running title: Expression pattern of selected histone demethylases in human granulosa cells","PeriodicalId":18329,"journal":{"name":"Medical Journal of Cell Biology","volume":"9 1","pages":"1 - 7"},"PeriodicalIF":0.0,"publicationDate":"2021-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41787418","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}
Abstract The awareness of the widespread influence of hypertension on various organ systems is ever increasing. Changes associated with this disease can be observed in the heart, brain, kidneys, but also the organ of vision. These usual microvascular changes are defined as hypertensive retinopathy. During a funduscopic examination, abnormalities such as narrowing of arterioles, symptoms of arteriole and vein intersection, cotton wool spots, intra-retinal exudates, retinal haemorrhages, and in severe cases even swelling of the optic disc and macula. This review presents an overview of the changes at the fundus of the eye that may occur in patients with hypertension, as well as problems with the classification of hypertensive retinopathy over the years, and the development of diagnostic methods in ophthalmology and fundoscopic imaging. Running title: The history of hypertensive retinopathy research
{"title":"The history of research of the ophthalmic aspects of hypertension","authors":"Aleksandra Krasińska, A. Brązert, J. Kocięcki","doi":"10.2478/acb-2021-0003","DOIUrl":"https://doi.org/10.2478/acb-2021-0003","url":null,"abstract":"Abstract The awareness of the widespread influence of hypertension on various organ systems is ever increasing. Changes associated with this disease can be observed in the heart, brain, kidneys, but also the organ of vision. These usual microvascular changes are defined as hypertensive retinopathy. During a funduscopic examination, abnormalities such as narrowing of arterioles, symptoms of arteriole and vein intersection, cotton wool spots, intra-retinal exudates, retinal haemorrhages, and in severe cases even swelling of the optic disc and macula. This review presents an overview of the changes at the fundus of the eye that may occur in patients with hypertension, as well as problems with the classification of hypertensive retinopathy over the years, and the development of diagnostic methods in ophthalmology and fundoscopic imaging. Running title: The history of hypertensive retinopathy research","PeriodicalId":18329,"journal":{"name":"Medical Journal of Cell Biology","volume":"9 1","pages":"14 - 18"},"PeriodicalIF":0.0,"publicationDate":"2021-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46589059","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}
M. Nawrocki, Rut Bryl, S. Kałużna, K. Stefańska, B. Stelmach, M. Jemielity, B. Perek, D. Bukowska, P. Mozdziak, J. Petitte, B. Kempisty
Abstract Heart failure (HF) is one of the main causes of death worldwide. Recent studies reported altered DNA methylation in failing human hearts. This may suggest a role of DNA methylation, most frequently implicated in epigenetic control, in the development of heart failure. Here, employing RT-qPCR, we characterized transcript levels for main DNA methyltransferases (DNMTs), DNMT1, DNMT3A, and DNMT3B, mediate DNA methylation, and they have different functions that complement each other during methylation. All analyzes were performed at different stages of porcine myocardial cell primary culture. In the present study we demonstrated increasing transcript expression levels for all analyzed genes during in vitro cultivation. The changes for DNMT1 and DNMT3A seem to be particularly important, where statistically significant changes were observed. Running title: DNMTs role in cardiac muscle cell culture
{"title":"Increased transcript expression levels of DNA methyltransferases type 1 and 3A during cardiac muscle long-term cell culture","authors":"M. Nawrocki, Rut Bryl, S. Kałużna, K. Stefańska, B. Stelmach, M. Jemielity, B. Perek, D. Bukowska, P. Mozdziak, J. Petitte, B. Kempisty","doi":"10.2478/acb-2021-0005","DOIUrl":"https://doi.org/10.2478/acb-2021-0005","url":null,"abstract":"Abstract Heart failure (HF) is one of the main causes of death worldwide. Recent studies reported altered DNA methylation in failing human hearts. This may suggest a role of DNA methylation, most frequently implicated in epigenetic control, in the development of heart failure. Here, employing RT-qPCR, we characterized transcript levels for main DNA methyltransferases (DNMTs), DNMT1, DNMT3A, and DNMT3B, mediate DNA methylation, and they have different functions that complement each other during methylation. All analyzes were performed at different stages of porcine myocardial cell primary culture. In the present study we demonstrated increasing transcript expression levels for all analyzed genes during in vitro cultivation. The changes for DNMT1 and DNMT3A seem to be particularly important, where statistically significant changes were observed. Running title: DNMTs role in cardiac muscle cell culture","PeriodicalId":18329,"journal":{"name":"Medical Journal of Cell Biology","volume":"9 1","pages":"27 - 32"},"PeriodicalIF":0.0,"publicationDate":"2021-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43928163","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}
Rut Bryl, C. Dompe, M. Jankowski, K. Stefańska, A. G. Narenji, Jakub Kulus, M. Kulus, M. Wieczorkiewicz, G. Wąsiatycz, J. Jaśkowski, M. Kaczmarek, J. Petitte, P. Mozdziak, P. Antosik, D. Bukowska
Abstract ADSCs are readily accessible and widely available. Isolated through a minimally invasive procedure from adipose depots, they can be found at diverse body location, where they served various functions, including energy homeostasis. They can be obtained upon surgeries from otherwise waste tissues, like after excision of fat tissue or liposuction. In addition, due to the possibility to isolate many ADSCs, in vitro proliferation can be performed in a short time period, resulting in cells showing more predictable results[1]. For this study ADSCs were obtained from waste material following routing sterilization procedures of dogs. This study aimed to analyse the expression of MSC specific markers before and after in vitro differentiation of ASCs. Three positive and three negative markers were analysed, CD105, CD73, CD90, CD34, CD14 and CD45. There were significant differences detected in the expression of all of the genes, with most of them exhibiting notable downregulation. The only exception, CD14 showed major upregulation after the process of differentiation. These changes confirm the success of differentiation, as well as suggest that this process significantly lowers the stem-like ability of ASCs. This knowledge should serve as a reference for further molecular and clinical studies, possibly aiding the understanding of the internal mechanisms governing the differentiation and stemness of ASCs, to enable their widespread and safe application in regenerative medicine. Running title: Mesenchymal markers during ASC osteogenic differentiation
{"title":"The influence of osteogenic differentiation on the stem-like properties of adipose derived stem cells – an RT-qPCR study","authors":"Rut Bryl, C. Dompe, M. Jankowski, K. Stefańska, A. G. Narenji, Jakub Kulus, M. Kulus, M. Wieczorkiewicz, G. Wąsiatycz, J. Jaśkowski, M. Kaczmarek, J. Petitte, P. Mozdziak, P. Antosik, D. Bukowska","doi":"10.2478/acb-2020-0020","DOIUrl":"https://doi.org/10.2478/acb-2020-0020","url":null,"abstract":"Abstract ADSCs are readily accessible and widely available. Isolated through a minimally invasive procedure from adipose depots, they can be found at diverse body location, where they served various functions, including energy homeostasis. They can be obtained upon surgeries from otherwise waste tissues, like after excision of fat tissue or liposuction. In addition, due to the possibility to isolate many ADSCs, in vitro proliferation can be performed in a short time period, resulting in cells showing more predictable results[1]. For this study ADSCs were obtained from waste material following routing sterilization procedures of dogs. This study aimed to analyse the expression of MSC specific markers before and after in vitro differentiation of ASCs. Three positive and three negative markers were analysed, CD105, CD73, CD90, CD34, CD14 and CD45. There were significant differences detected in the expression of all of the genes, with most of them exhibiting notable downregulation. The only exception, CD14 showed major upregulation after the process of differentiation. These changes confirm the success of differentiation, as well as suggest that this process significantly lowers the stem-like ability of ASCs. This knowledge should serve as a reference for further molecular and clinical studies, possibly aiding the understanding of the internal mechanisms governing the differentiation and stemness of ASCs, to enable their widespread and safe application in regenerative medicine. Running title: Mesenchymal markers during ASC osteogenic differentiation","PeriodicalId":18329,"journal":{"name":"Medical Journal of Cell Biology","volume":"8 1","pages":"158 - 163"},"PeriodicalIF":0.0,"publicationDate":"2020-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45531466","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}
S. Kałużna, Rut Bryl, B. Chermuła, Rafał Sibiak, K. Stefańska, W. Pieńkowski, W. Kranc, M. Ješeta, P. Ventruba, J. Žáková, I. Crha, L. Pawelczyk, R. Spaczyński
Abstract The essential function of granulosa cells is to maintain the proper course of oogenesis and folliculogenesis. The immune system is an additional local regulator of ovarian function, with cytokines necessary for the proper function of the ovaries, including the secretion of steroid hormones This study aimed to analyze the expression of genes in human GCs in short-term primary culture and define the difference in the expression of IL1β, IL6, and TNFα genes at 48h and 72h of culture compared to the 24h control. Total RNA was isolated using the Chomczyński and Sacchi protocol. RNA samples were treated with DNase I and reverse transcribed (RT) into cDNA. The determination of transcript levels of the mentioned genes was performed using the Light Cycler® 96 Real-Time PCR kit, Roche Diagnostics GmbH (Mannheim, Germany). The present study proved that granulosa cells in a short-term primary in vitro culture express IL-1β, IL-6, and TNFα. The tested genes show a decrease in expression at 24h of culture and a subsequent slight increase at 72h, not exceeding the initial levels. The expression changes the most for IL1β and the least for TNFα. The fluctuations in the amount of transcript may be influenced by factors stored in granulosa cells before the IVM procedure, the procedure of in vitro fertilization, as well as factors related to the process of primary culture. More research is needed to understand the details of these occurrences. Running title: The inflammatory response in human granulosa cells
{"title":"Expression of genes involved in the inflammatory response in human granulosa cells in short-term in vitro culture","authors":"S. Kałużna, Rut Bryl, B. Chermuła, Rafał Sibiak, K. Stefańska, W. Pieńkowski, W. Kranc, M. Ješeta, P. Ventruba, J. Žáková, I. Crha, L. Pawelczyk, R. Spaczyński","doi":"10.2478/acb-2020-0025","DOIUrl":"https://doi.org/10.2478/acb-2020-0025","url":null,"abstract":"Abstract The essential function of granulosa cells is to maintain the proper course of oogenesis and folliculogenesis. The immune system is an additional local regulator of ovarian function, with cytokines necessary for the proper function of the ovaries, including the secretion of steroid hormones This study aimed to analyze the expression of genes in human GCs in short-term primary culture and define the difference in the expression of IL1β, IL6, and TNFα genes at 48h and 72h of culture compared to the 24h control. Total RNA was isolated using the Chomczyński and Sacchi protocol. RNA samples were treated with DNase I and reverse transcribed (RT) into cDNA. The determination of transcript levels of the mentioned genes was performed using the Light Cycler® 96 Real-Time PCR kit, Roche Diagnostics GmbH (Mannheim, Germany). The present study proved that granulosa cells in a short-term primary in vitro culture express IL-1β, IL-6, and TNFα. The tested genes show a decrease in expression at 24h of culture and a subsequent slight increase at 72h, not exceeding the initial levels. The expression changes the most for IL1β and the least for TNFα. The fluctuations in the amount of transcript may be influenced by factors stored in granulosa cells before the IVM procedure, the procedure of in vitro fertilization, as well as factors related to the process of primary culture. More research is needed to understand the details of these occurrences. Running title: The inflammatory response in human granulosa cells","PeriodicalId":18329,"journal":{"name":"Medical Journal of Cell Biology","volume":"8 1","pages":"190 - 195"},"PeriodicalIF":0.0,"publicationDate":"2020-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42724381","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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