Nan Zhang, Li Xu, Hao Song, Chunqing Bu, Jie Kang, Chuanchen Zhang, Xiaofei Yang, Fabin Han
{"title":"Tracking of Stem Cells from Human Exfoliated Deciduous Teeth Labeled with Molday ION Rhodamine-B during Periodontal Bone Regeneration in Rats.","authors":"Nan Zhang, Li Xu, Hao Song, Chunqing Bu, Jie Kang, Chuanchen Zhang, Xiaofei Yang, Fabin Han","doi":"10.15283/ijsc21204","DOIUrl":"https://doi.org/10.15283/ijsc21204","url":null,"abstract":"<p><strong>Background and objectives: </strong>Chronic periodontitis can lead to alveolar bone resorption and eventually tooth loss. Stem cells from exfoliated deciduous teeth (SHED) are appropriate bone regeneration seed cells. To track the survival, migration, and differentiation of the transplanted SHED, we used super paramagnetic iron oxide particles (SPIO) Molday ION Rhodamine-B (MIRB) to label and monitor the transplanted cells while repairing periodontal bone defects.</p><p><strong>Methods and results: </strong>We determined an appropriate dose of MIRB for labeling SHED by examining the growth and osteogenic differentiation of labeled SHED. Finally, SHED was labeled with 25 μg Fe/ml MIRB before being transplanted into rats. Magnetic resonance imaging was used to track SHED survival and migration <i>in vivo</i> due to a low-intensity signal artifact caused by MIRB. HE and immunohistochemical analyses revealed that both MIRB-labeled and unlabeled SHED could promote periodontal bone regeneration. The colocalization of hNUC and MIRB demonstrated that SHED transplanted into rats could survive <i>in vivo</i>. Furthermore, some MIRB-positive cells expressed the osteoblast and osteocyte markers OCN and DMP1, respectively. Enzyme-linked immunosorbent assay revealed that SHED could secrete protein factors, such as IGF-1, OCN, ALP, IL-4, VEGF, and bFGF, which promote bone regeneration. Immunofluorescence staining revealed that the transplanted SHED was surrounded by a large number of host-derived Runx2- and Col II-positive cells that played important roles in the bone healing process.</p><p><strong>Conclusions: </strong>SHED could promote periodontal bone regeneration in rats, and the survival of SHED could be tracked <i>in vivo</i> by labeling them with MIRB. SHED are likely to promote bone healing through both direct differentiation and paracrine mechanisms.</p>","PeriodicalId":14392,"journal":{"name":"International journal of stem cells","volume":"16 1","pages":"93-107"},"PeriodicalIF":2.3,"publicationDate":"2023-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/9f/6e/ijsc-16-1-93.PMC9978830.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9386474","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Wenyong Fei, Erkai Pang, Lei Hou, Jihang Dai, Mingsheng Liu, Xuanqi Wang, Bin Xie, Jingcheng Wang
{"title":"Synergistic Effect of Hydrogen and 5-Aza on Myogenic Differentiation through the p38 MAPK Signaling Pathway in Adipose-Derived Mesenchymal Stem Cells.","authors":"Wenyong Fei, Erkai Pang, Lei Hou, Jihang Dai, Mingsheng Liu, Xuanqi Wang, Bin Xie, Jingcheng Wang","doi":"10.15283/ijsc21238","DOIUrl":"https://doi.org/10.15283/ijsc21238","url":null,"abstract":"<p><strong>Background and objectives: </strong>This study aims to clarify the systems underlying regulation and regulatory roles of hydrogen combined with 5-Aza in the myogenic differentiation of adipose mesenchymal stem cells (ADSCs).</p><p><strong>Methods and results: </strong>In this study, ADSCs acted as an in vitro myogenic differentiating mode. First, the Alamar blue Staining and mitochondrial tracer technique were used to verify whether hydrogen combined with 5-Aza could promote cell proliferation. In addition, this study assessed myogenic differentiating markers (e.g., Myogenin, Mhc and Myod protein expressions) based on the Western blotting assay, analysis on cellular morphological characteristics (e.g., Myotube number, length, diameter and maturation index), RT-PCR (Myod, Myogenin and Mhc mRNA expression) and Immunofluorescence analysis (Desmin, Myosin and β-actin protein expression). Finally, to verify the mechanism of myogenic differentiation of hydrogen-bound 5-Aza, we performed bioinformatics analysis and Western blot to detect the expression of p-P38 protein. Hydrogen combined with 5-Aza significantly enhanced the proliferation and myogenic differentiation of ADSCs in vitro by increasing the number of single-cell mitochondria and upregulating the expression of myogenic biomarkers such as Myod, Mhc and myotube formation. The expressions of p-P38 was up-regulated by hydrogen combined with 5-Aza. The differentiating ability was suppressed when the cells were cultivated in combination with SB203580 (p38 MAPK signal pathway inhibitor).</p><p><strong>Conclusions: </strong>Hydrogen alleviates the cytotoxicity of 5-Aza and synergistically promotes the myogenic differentiation capacity of adipose stem cells via the p38 MAPK pathway. Thus, the mentioned results present insights into myogenic differentiation and are likely to generate one potential alternative strategy for skeletal muscle related diseases.</p>","PeriodicalId":14392,"journal":{"name":"International journal of stem cells","volume":"16 1","pages":"78-92"},"PeriodicalIF":2.3,"publicationDate":"2023-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/10/53/ijsc-16-1-78.PMC9978834.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9370096","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Bong Jong Seo, Tae Kyung Hong, Sang Hoon Yoon, Jae Hoon Song, Sang Jun Uhm, Hyuk Song, Kwonho Hong, Hans Robert Schöler, Jeong Tae Do
{"title":"Embryonic Stem Cells Lacking DNA Methyltransferases Differentiate into Neural Stem Cells that Are Defective in Self-Renewal.","authors":"Bong Jong Seo, Tae Kyung Hong, Sang Hoon Yoon, Jae Hoon Song, Sang Jun Uhm, Hyuk Song, Kwonho Hong, Hans Robert Schöler, Jeong Tae Do","doi":"10.15283/ijsc22138","DOIUrl":"https://doi.org/10.15283/ijsc22138","url":null,"abstract":"<p><strong>Background and objectives: </strong>DNA methyltransferases (Dnmts) play an important role in regulating DNA methylation during early developmental processes and cellular differentiation. In this study, we aimed to investigate the role of Dnmts in neural differentiation of embryonic stem cells (ESCs) and in maintenance of the resulting neural stem cells (NSCs).</p><p><strong>Methods and results: </strong>We used three types of Dnmt knockout (KO) ESCs, including Dnmt1 KO, Dnmt3a/3b double KO (Dnmt3 DKO), and Dnmt1/3a/3b triple KO (Dnmt TKO), to investigate the role of Dnmts in neural differentiation of ESCs. All three types of Dnmt KO ESCs could form neural rosette and differentiate into NSCs <i>in vitro</i>. Interestingly, however, after passage three, Dnmt KO ESC-derived NSCs could not maintain their self-renewal and differentiated into neurons and glial cells.</p><p><strong>Conclusions: </strong>Taken together, the data suggested that, although deficiency of Dnmts had no effect on the differentiation of ESCs into NSCs, the latter had defective maintenance, thereby indicating that Dnmts are crucial for self-renewal of NSCs.</p>","PeriodicalId":14392,"journal":{"name":"International journal of stem cells","volume":"16 1","pages":"44-51"},"PeriodicalIF":2.3,"publicationDate":"2023-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/5f/56/ijsc-16-1-44.PMC9978838.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10831200","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Yu Zhao, Chuan Guo, Lianlin Zeng, Jialing Li, Xia Liu, Yiwei Wang, Kun Zhao, Bo Chen
{"title":"Mesenchymal Stem Cells Ameliorate Fibrosis by Enhancing Autophagy via Inhibiting Galectin-3/Akt/mTOR Pathway and by Alleviating the EMT via Inhibiting Galectin-3/Akt/GSK3β/Snail Pathway in NRK-52E Fibrosis.","authors":"Yu Zhao, Chuan Guo, Lianlin Zeng, Jialing Li, Xia Liu, Yiwei Wang, Kun Zhao, Bo Chen","doi":"10.15283/ijsc22014","DOIUrl":"https://doi.org/10.15283/ijsc22014","url":null,"abstract":"<p><strong>Background and objectives: </strong>Epithelial-Mesenchymal transition (EMT) is one of the origins of myofibroblasts in renal interstitial fibrosis. Mesenchymal stem cells (MSCs) alleviating EMT has been proved, but the concrete mechanism is unclear. To explore the mechanism, serum-free MSCs conditioned medium (SF-MSCs-CM) was used to treat rat renal tubular epithelial cells (NRK-52E) fibrosis induced by transforming growth factor-β1 (TGF-β1) which ameliorated EMT.</p><p><strong>Methods and results: </strong>Galectin-3 knockdown (Gal-3 KD) and overexpression (Gal-3 OE) lentiviral vectors were established and transfected into NRK-52E. NRK-52E fibrosis model was induced by TGF-β1 and treated with the SF-MSCs-CM for 24 h after modelling. Fibrosis and autophagy related indexes were detected by western blot and immunocytochemistry. In model group, the expressions of α-smooth muscle actin (α-SMA), fibronectin (FN), Galectin-3, Snail, Kim-1, and the ratios of P-Akt/Akt, P-GSK3β/GSK3β, P-PI3K/PI3K, P-mTOR/mTOR, TIMP1/MMP9, and LC3B-II/I were obviously increased, and E-Cadherin (E-cad) and P62 decreased significantly compared with control group. SF-MSCs-CM showed an opposite trend after treatment compared with model group. Whether in Gal-3 KD or Gal-3 OE NRK-52E cells, SF-MSCs-CM also showed similar trends. However, the effects of anti-fibrosis and enhanced autophagy in Gal-3 KD cells were more obvious than those in Gal-3 OE cells.</p><p><strong>Conclusions: </strong>SF-MSCs-CM probably alleviated the EMT via inhibiting Galectin-3/Akt/GSK3β/Snail pathway. Meanwhile, Gal-3 KD possibly enhanced autophagy via inhibiting Galectin-3/Akt/mTOR pathway, which synergistically ameliorated renal fibrosis. Targeting galectin-3 may be a potential target for the treatment of renal fibrosis.</p>","PeriodicalId":14392,"journal":{"name":"International journal of stem cells","volume":"16 1","pages":"52-65"},"PeriodicalIF":2.3,"publicationDate":"2023-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/84/04/ijsc-16-1-52.PMC9978829.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9386448","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Myunghyun Kim, Junmyeong Park, Sujin Kim, Dong Wook Han, Borami Shin, Hans Robert Schöler, Johnny Kim, Kee-Pyo Kim
{"title":"Generation of Induced Pluripotent Stem Cells from Lymphoblastoid Cell Lines by Electroporation of Episomal Vectors.","authors":"Myunghyun Kim, Junmyeong Park, Sujin Kim, Dong Wook Han, Borami Shin, Hans Robert Schöler, Johnny Kim, Kee-Pyo Kim","doi":"10.15283/ijsc22177","DOIUrl":"https://doi.org/10.15283/ijsc22177","url":null,"abstract":"Background and Objectives Lymphoblastoid cell lines (LCLs) deposited from disease-affected individuals could be a valuable donor cell source for generating disease-specific induced pluripotent stem cells (iPSCs). However, generation of iPSCs from the LCLs is still challenging, as yet no effective gene delivery strategy has been developed. Methods and Results Here, we reveal an effective gene delivery method specifically for LCLs. We found that LCLs appear to be refractory toward retroviral and lentiviral transduction. Consequently, lentiviral and retroviral transduction of OCT4, SOX2, KFL4 and c-MYC into LCLs does not elicit iPSC colony formation. Interestingly, however we found that transfection of oriP/EBNA-1-based episomal vectors by electroporation is an efficient gene delivery system into LCLs, enabling iPSC generation from LCLs. These iPSCs expressed pluripotency makers (OCT4, NANOG, SSEA4, SALL4) and could form embryoid bodies. Conclusions Our data show that electroporation is an effective gene delivery method with which LCLs can be efficiently reprogrammed into iPSCs.","PeriodicalId":14392,"journal":{"name":"International journal of stem cells","volume":"16 1","pages":"36-43"},"PeriodicalIF":2.3,"publicationDate":"2023-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/ad/e9/ijsc-16-1-36.PMC9978833.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10822360","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Yukyeong Lee, Seung-Won Lee, Dahee Jeong, Hye Jeong Lee, Na Young Choi, Jin Seok Bang, Seokbeom Ham, Kinarm Ko
{"title":"Inhibition of Class I Histone Deacetylase Enhances Self-Reprogramming of Spermatogonial Stem Cells into Pluripotent Stem Cells.","authors":"Yukyeong Lee, Seung-Won Lee, Dahee Jeong, Hye Jeong Lee, Na Young Choi, Jin Seok Bang, Seokbeom Ham, Kinarm Ko","doi":"10.15283/ijsc22110","DOIUrl":"https://doi.org/10.15283/ijsc22110","url":null,"abstract":"<p><strong>Background and objectives: </strong>Spermatogonial stem cells (SSCs) are the most primitive cells in spermatogenesis and are the only adult stem cells capable of passing on the genome of a given species to the next generation. SSCs are the only adult stem cells known to exhibit high Oct4 expression and can be induced to self-reprogram into pluripotent cells depending on culture conditions. Epigenetic modulation is well known to be involved in the induction of pluripotency of somatic cells. However, epigenetic modulation in self-reprogramming of SSCs into pluripotent cells has not been studied.</p><p><strong>Methods and results: </strong>In this study, we examined the involvement of epigenetic modulation by assessing whether self-reprogramming of SSCs is enhanced by treatment with epigenetic modulators. We found that second-generation selective class I HDAC inhibitors increased SSC reprogramming efficiency, whereas non-selective HDAC inhibitors had no effect.</p><p><strong>Conclusions: </strong>We showed that pluripotent stem cells derived from adult SSCs by treatment with small molecules with epigenetic modulator functions exhibit pluripotency <i>in vitro</i> and <i>in vivo</i>. Our results suggest that the mechanism of SSC reprogramming by epigenetic modulator can be used for important applications in epigenetic reprogramming research.</p>","PeriodicalId":14392,"journal":{"name":"International journal of stem cells","volume":"16 1","pages":"27-35"},"PeriodicalIF":2.3,"publicationDate":"2023-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/7e/0f/ijsc-16-1-27.PMC9978831.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10831653","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Hyo Jin Kim, Gyeongmin Kim, Kyun Yoo Chi, Jong-Hoon Kim
{"title":"<i>In Vitro</i> Generation of Luminal Vasculature in Liver Organoids: From Basic Vascular Biology to Vascularized Hepatic Organoids.","authors":"Hyo Jin Kim, Gyeongmin Kim, Kyun Yoo Chi, Jong-Hoon Kim","doi":"10.15283/ijsc22154","DOIUrl":"https://doi.org/10.15283/ijsc22154","url":null,"abstract":"<p><p>Liver organoids have gained much attention in recent years for their potential applications to liver disease modeling and pharmacologic drug screening. Liver organoids produced <i>in vitro</i> reflect some aspects of the in vivo physiological and pathological conditions of the liver. However, the generation of liver organoids with perfusable luminal vasculature remains a major challenge, hindering precise and effective modeling of liver diseases. Furthermore, vascularization is required for large organoids or assembloids to closely mimic the complexity of tissue architecture without cell death in the core region. A few studies have successfully generated liver organoids with endothelial cell networks, but most of these vascular networks produced luminal structures after being transplanted into tissues of host animals. Therefore, formation of luminal vasculature is an unmet need to overcome the limitation of liver organoids as an <i>in vitro</i> model investigating different acute and chronic liver diseases. Here, we provide an overview of the unique features of hepatic vasculature under pathophysiological conditions and summarize the biochemical and biophysical cues that drive vasculogenesis and angiogenesis <i>in vitro</i>. We also highlight recent progress in generating vascularized liver organoids <i>in vitro</i> and discuss potential strategies that may enable the generation of perfusable luminal vasculature in liver organoids.</p>","PeriodicalId":14392,"journal":{"name":"International journal of stem cells","volume":"16 1","pages":"1-15"},"PeriodicalIF":2.3,"publicationDate":"2023-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/dd/c1/ijsc-16-1-1.PMC9978835.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10831203","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Han Kyu Chae, Nayoung Suh, Myong Jin Jang, Yu Seon Kim, Bo Hyun Kim, Joomin Aum, Ha Chul Shin, Dalsan You, Bumsik Hong, Hyung Keun Park, Choung-Soo Kim
{"title":"Efficacy and Safety of Human Bone Marrow-Derived Mesenchymal Stem Cells according to Injection Route and Dose in a Chronic Kidney Disease Rat Model.","authors":"Han Kyu Chae, Nayoung Suh, Myong Jin Jang, Yu Seon Kim, Bo Hyun Kim, Joomin Aum, Ha Chul Shin, Dalsan You, Bumsik Hong, Hyung Keun Park, Choung-Soo Kim","doi":"10.15283/ijsc21146","DOIUrl":"https://doi.org/10.15283/ijsc21146","url":null,"abstract":"<p><strong>Background and objectives: </strong>We compared the efficacy and safety of human bone marrow-derived mesenchymal stem cells (hBMSC), delivered at different doses and via different injection routes in an animal model of chronic kidney disease.</p><p><strong>Methods and results: </strong>A total of ninety 12-week-old rats underwent 5/6 nephrectomy and randomized among nine groups: sham, renal artery control (RA-C), tail vein control (TV-C), renal artery low dose (RA-LD) (0.5×10<sup>6</sup> cells), renal artery moderate dose (RA-MD) (1.0×10<sup>6</sup> cells), renal artery high dose (RA-HD) (2.0×10<sup>6</sup> cells), tail vein low dose (TV-LD) (0.5×10<sup>6</sup> cells), tail vein moderate dose (TV-MD) (1.0×10<sup>6</sup> cells), and tail vein high dose (TV-HD) (2.0×10<sup>6</sup> cells). Renal function and mortality of rats were evaluated after hBMSC injection. Serum blood urea nitrogen was significantly lower in the TV-HD group at 2 weeks (p<0.01), 16 weeks (p<0.05), and 24 weeks (p<0.01) than in the TV-C group, as determined by one-way ANOVA. Serum creatinine was significantly lower in the TV-HD group at 24 weeks (p<0.05). At 8 weeks, creatinine clearance was significantly higher in the TV-MD and TV-HD groups (p<0.01, p<0.05) than in the TV-C group. In the safety evaluation, we observed no significant difference among the groups.</p><p><strong>Conclusions: </strong>Our findings confirm the efficacy and safety of high dose (2×10<sup>6</sup> cells) injection of hBMSC via the tail vein.</p>","PeriodicalId":14392,"journal":{"name":"International journal of stem cells","volume":"16 1","pages":"66-77"},"PeriodicalIF":2.3,"publicationDate":"2023-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/b9/ca/ijsc-16-1-66.PMC9978839.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9386447","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Soo-Been Jeon, A-Reum Han, Yoo Bin Choi, Ah Reum Lee, Ji Yoon Lee
{"title":"Lymphoid Lineage γδ T Cells Were Successfully Generated from Human Pluripotent Stem Cells via Hemogenic Endothelium.","authors":"Soo-Been Jeon, A-Reum Han, Yoo Bin Choi, Ah Reum Lee, Ji Yoon Lee","doi":"10.15283/ijsc22150","DOIUrl":"https://doi.org/10.15283/ijsc22150","url":null,"abstract":"<p><p>γδ T cells are a rare and unique prototype of T cells that share properties with natural killer cells in secondary lymphoid organs. Although many studies have revealed the function and importance of adult-derived γδ T cells in cancer biology and regenerative medicine, the low numbers of these cells hamper their application as therapeutic cell sources in the clinic. To solve this problem, pluripotent stem cell-derived γδ T cells are considered alternative cell sources; however, few studies have reported the generation of human pluripotent stem cell-derived γδ T cells. In the present study, we investigated whether lymphoid lineage γδ T cells were successfully generated from human pluripotent stem cells via hemogenic endothelium under defined culture conditions. Our results revealed that pluripotent stem cells successfully generated γδ T cells with an overall increase in transcriptional activity of lymphoid lineage genes and cytolytic factors, indicating the importance of the optimization of culture conditions in generating lymphoid lineage γδ T cells. We uncovered an initial step in differentiating γδ T cells that could be applied to basic and translational investigations in the field of cancer biology. Based on our result, we will develop an appropriate method to purify γδ T cells with functionality and it helpful for the study of basic mechanism of γδ T cells in pathophysiologic condition as well as clinic application.</p>","PeriodicalId":14392,"journal":{"name":"International journal of stem cells","volume":"16 1","pages":"108-116"},"PeriodicalIF":2.3,"publicationDate":"2023-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/4e/3f/ijsc-16-1-108.PMC9978832.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10831202","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Seung Hwan Ko, Jin Sun Kang, Sang-Mi Kim, Eun-Hye Lee, Chang-Hwan Park
{"title":"Reduced Cytotoxicity by Repetitive mRNA Transfection in Differentiated Neurons.","authors":"Seung Hwan Ko, Jin Sun Kang, Sang-Mi Kim, Eun-Hye Lee, Chang-Hwan Park","doi":"10.15283/ijsc22125","DOIUrl":"https://doi.org/10.15283/ijsc22125","url":null,"abstract":"<p><strong>Background and objectives: </strong>mRNA-based protein expression technology has been used to express functional proteins. We have previously generated dopamine neurons from rat-embryo derived neural precursor cells (NPCs) through repeated transfection of synthetic transcription factor mRNA encoding dopamine-inducible genes. However, NPCs began to die approximately 10 d post-transfection. In this study, we examined a long-term transfection protocol that did not affect cell viability.</p><p><strong>Methods and results: </strong>Experiments were performed in eight groups sorted according to the start date of mRNA transfection. mRNA was transfected into NPCs daily for 21 d and live cell images of each group were recorded. NPCs which were differentiated for more than five days showed sustained gene expression and appreciable viability despite daily mRNA transfection for 21 d.</p><p><strong>Conclusions: </strong>Repeated mRNA transfection requires cells with a sufficient differentiation period.</p>","PeriodicalId":14392,"journal":{"name":"International journal of stem cells","volume":"16 1","pages":"117-122"},"PeriodicalIF":2.3,"publicationDate":"2023-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/a1/83/ijsc-16-1-117.PMC9978836.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10831652","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}