Heterogeneity of pluripotent marker gene expression in colonies generated in human iPS cell induction culture

Hara, Masaki; Ishikawa, Tetsuya; Takahashi, S.; Okumura, Mitsutaka; Sakai, Noriko; Haga, Megumi; Kominami, Katsuya; Migita, Hideyuki; McDonald, Fiona; Shimada, Fumiki; Sakurada, Kazuhiro

doi:10.1016/j.scr.2008.01.001

Cited by 71 publications

(75 citation statements)

References 30 publications

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“…BJ1, BJ2, and BJ fibroblasts are three different foreskin lines containing the telomerase gene; while FS is a foreskin fibroblast primary cell line, HIF indicates human embryonic fibroblasts; Derm indicates human neonatal dermal fibroblasts; MRC5 are primary fetal cells. The hESC lines where HI-OGN, HUES 8, HSF1, H9, and HESC H9 [11][12][13][14]. The BJ1 cell line was originally designated BJ48 and its iPS cell line is BJ-iPS1.…”

Section: Discussionmentioning

confidence: 99%

Reprogramming of the MHC‐I and Its Regulation by NFκB in Human‐Induced Pluripotent Stem Cells

et al. 2012

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The immunogenicity of human pluripotent stem cells plays a major role in their potential use in the clinic. We show that, during their reprogramming, human-induced pluripotent stem (iPS) cells downregulate expression of human leukocyte antigen (HLA)-A/B/C and b2 microglobulin (b2M), the two components of major histocompatibility complex-I (MHC-I). MHC-I expression in iPS cells can be restored by differentiation or treatment with interferongamma (IFNc). To analyze the molecular mechanisms that regulate the expression of the MHC-I molecules in human iPS cells, we searched for correlation between the expression of HLA-A/B/C and b2M and the expression of transcription factors that bind to the promoter of these genes. Our results show a significant positive correlation between MHC-I expression and expression of the nuclear factors, nuclear factor kappa B 1 (NFjB1) and RelA, at the levels of RNA, protein and was confirmed by chromatin binding. Concordantly, we detected robust levels of NFjB1 and RelA proteins in the nucleus of somatic cells but not in the iPS cell derived from them. Overexpression of NFjB1 and RelA in undifferentiated pluripotent stem cells led to induction in expression of MHC-I, whereas silencing NFjB1 and RelA by small hairpin RNA decreased the expression of b2M after IFNc treatment. Our data point to the critical role of NFjB proteins in regulating the MHC-I expression in human pluripotent stem cells. STEM

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Section: Discussionmentioning

confidence: 99%

Reprogramming of the MHC‐I and Its Regulation by NFκB in Human‐Induced Pluripotent Stem Cells

et al. 2012

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“…One of the critical problems is absence of ready-to-use stem cell sources. Embryonic stem cells [1] or induced pluripotent stem cells [2] have cardiomyogenic differentiation potential and are major candidates for future stem cell therapy, but, there are still several problems to overcome before clinical application, that is, neoplasm formation [3] and chromosomal stability [4]. Human bone marrow-derived mesenchymal stem cells (BM-MSCs) have a cardiomyogenic transdifferentiation ability; however, the cardiomyogenic transdifferentiation efficiency (CTE) is extremely low [5].…”

Section: Introductionmentioning

confidence: 99%

Pretreatment of Human Mesenchymal Stem Cells with Pioglitazone Improved Efficiency of Cardiomyogenic Transdifferentiation and Cardiac Function

et al. 2011

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The efficacy of transplantation of default human marrowderived mesenchymal stem cells (MSCs) was modest. In this study, our challenge was to improve the efficacy of MSC transplantation in vivo by pretreatment of MSCs with pioglitazone. MSCs were cultured with or without medium containing 1 lM of pioglitazone before cardiomyogenic induction. After cardiomyogenic induction in vitro, cardiomyogenic transdifferentiation efficiency (CTE) was calculated by immunocytochemistry using anti-cardiac troponin-I antibody. For the in vivo experiments, myocardial infarction (MI) at the anterior left ventricle was made in nude rats. Two weeks after MI, MSCs pretreated with pioglitazone (p-BM; n 5 30) or without pioglitazone (BM; n 5 17) were injected, and then survived for 2 weeks. We compared left ventricular function by echocardiogram and immunohistochemistry to observe cardiomyogenic transdifferentiation in vivo. Pretreatment with pioglitazone significantly increased the CTE in vitro (1.9% 6 0.2% n 5 47 vs. 39.5% 6 4.7% n 5 13, p < .05). Transplantation of pioglitazone pretreated MSCs significantly improved change in left ventricular % fractional shortening (BM; 24.8% 6 2.1%, vs. p-BM; 5.2% 6 1.5%). Immunohistochemistry revealed significant improvement of cardiomyogenic transdifferentiation in p-BM in vivo (BM; 0% 6 0% n 5 5, vs. p-BM; 0.077% 6 0.041% n 5 5). Transplantation of pioglitazonepretreated MSCs significantly improved cardiac function and can be a promising cardiac stem cell source to expect cardiomyogenesis.

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“…PCR was done with LATaq (Takara-Bio). Oligonucleotide primer sequences were previously described [6,20] and as follows: glyceraldehyde 3-phophate dehydrogenase forward, 5 0 -ACCACAGTCCATG CCATCAC -3 0 , and glyceraldehyde 3-phophate dehydrogenase reverse, 5 0 -TCCACCACCCTGTTGCTGTA -3 0 .…”

Section: Gene Expression Analysismentioning

confidence: 99%

Direct Generation of Induced Pluripotent Stem Cells from Human Nonmobilized Blood

Kunisato

Wakatsuki

Shinba

et al. 2011

Stem Cells and Development

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The use of induced pluripotent stem cells (iPSCs) is an exciting frontier in the study and treatment of human diseases through the generation of specific cell types. Here we show the derivation of iPSCs from human nonmobilized peripheral blood (PB) and bone marrow (BM) mononuclear cells (MNCs) by retroviral transduction of OCT3/4, SOX2, KLF4, and c-MYC. The PB- and BM-derived iPSCs were quite similar to human embryonic stem cells with regard to morphology, expression of surface antigens and pluripotency-associated transcription factors, global gene expression profiles, and differentiation potential in vitro and in vivo. Infected PB and BM MNCs gave rise to iPSCs in the presence of several cytokines, although transduction efficiencies were not high. We found that 5 × 10(5) PB MNCs, which corresponds to less than 1 mL of PB, was enough for the generation of several iPSC colonies. Generation of iPSCs from MNCs of nonmobilized PB, with its relative efficiency and ease of harvesting, could enable the therapeutic use of patient-specific pluripotent stem cells.

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Heterogeneity of pluripotent marker gene expression in colonies generated in human iPS cell induction culture

Cited by 71 publications

References 30 publications

Reprogramming of the MHC‐I and Its Regulation by NFκB in Human‐Induced Pluripotent Stem Cells

Reprogramming of the MHC‐I and Its Regulation by NFκB in Human‐Induced Pluripotent Stem Cells

Pretreatment of Human Mesenchymal Stem Cells with Pioglitazone Improved Efficiency of Cardiomyogenic Transdifferentiation and Cardiac Function

Direct Generation of Induced Pluripotent Stem Cells from Human Nonmobilized Blood

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