Monolayer endoderm differentiation from human ESCs

Cheng, Xin; Lei, Ying; Lin, Li; Galvão, André Luiz Baptista; Mills, JA; Hc, Lin; Kotton, D.N.; Shen, SS; Nostro, MC; Jk, Choi; Weiss, MJ; French, DL; Gadue, Paul

doi:10.3824/stembook.1.64.1

Cited by 2 publications

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“…When the hESCs reached 50% confluence, hepatic differentiation was induced by a stepwise strategy, as reported, with modification (Fig. 2A) [17]. hESCs were treated with 100 ng/ml activin A and 2 μM CHIR99021 for 2 days in insulin‐free differentiation medium to induce primitive streak, which was then patterned toward definitive endoderm by treatment with 100 ng/ml activin A, 10 ng/ml VEGF, 10 ng/ml FGF2, and 0.25 ng/ml BMP4 for another 3 days.…”

Section: Resultsmentioning

confidence: 95%

Generation of Self-Renewing Hepatoblasts From Human Embryonic Stem Cells by Chemical Approaches

Zhang

Sun

Wang

et al. 2015

Stem Cells Translational Medicine

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Somatic stem cells play crucial roles in organogenesis and tissue homeostasis and regeneration and may ultimately prove useful for cell therapy for a variety of degenerative diseases and injuries; however, isolation and expansion of most types of somatic stem cells from tissues are technically challenging. Human pluripotent stem cells are a renewable source for any adult cell types, including somatic stem cells. Generation of somatic stem cells from human pluripotent stem cells is a promising strategy to get these therapeutically valuable cells. Previously, we developed a chemically defined condition for mouse hepatoblast self-renewal through a reiterative screening strategy. In the present study, we efficiently generated hepatoblasts from human embryonic stem cells by a stepwise induction strategy. Importantly, these human embryonic stem cell-derived hepatoblasts can be captured and stably maintained using conditions previously established for mouse hepatoblast self-renewal, which includes basal media supplemented with insulin, transferrin, sodium selenite, epidermal growth factor, glycogen synthase kinase 3 inhibitor, transforming growth factor b receptor inhibitor, lysophosphatidic acid, and sphingosine 1-phosphate. The cells can stably retain hepatoblast phenotypes during prolonged culture and can differentiate into mature hepatocytes through in vitro provision of hepatocyte lineage developmental cues. After being embedded into three-dimensional Matrigel, these cells efficiently formed bile duct-like structures resembling native bile duct tissues. These human embryonic stem cell-derived hepatoblasts would be useful as a renewable source for cell therapy of liver diseases. STEM CELLS TRANSLATIONAL MEDICINE 2015;4:1275-1282 SIGNIFICANCE Somatic stem cells have been proposed as promising candidates for cell-based therapy; however, isolation of somatic stem cells from adult tissues is usually invasive and technically challenging. In the present study, hepatoblasts from human embryonic stem cells were efficiently generated. These human hepatoblasts were then stably captured and maintained by a growth factor and small molecule cocktail, which included epidermal growth factor, glycogen synthase kinase 3 inhibitor, transforming growth factor b receptor inhibitor, lysophosphatidic acid, and sphingosine 1-phosphate. These human embryonic stem cell-derived hepatoblasts would be useful as a renewable source for cell therapy of liver diseases.

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Section: Resultsmentioning

confidence: 95%

Generation of Self-Renewing Hepatoblasts From Human Embryonic Stem Cells by Chemical Approaches

Zhang

Sun

Wang

et al. 2015

Stem Cells Translational Medicine

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High-efficiency RNA-based reprogramming of human primary fibroblasts

et al. 2018

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Induced pluripotent stem cells (iPSCs) hold great promise for regenerative medicine; however, their potential clinical application is hampered by the low efficiency of somatic cell reprogramming. Here, we show that the synergistic activity of synthetic modified mRNAs encoding reprogramming factors and miRNA-367/302s delivered as mature miRNA mimics greatly enhances the reprogramming of human primary fibroblasts into iPSCs. This synergistic activity is dependent upon an optimal RNA transfection regimen and culturing conditions tailored specifically to human primary fibroblasts. As a result, we can now generate up to 4,019 iPSC colonies from only 500 starting human primary neonatal fibroblasts and reprogram up to 90.7% of individually plated cells, producing multiple sister colonies. This methodology consistently generates clinically relevant, integration-free iPSCs from a variety of human patient’s fibroblasts under feeder-free conditions and can be applicable for the clinical translation of iPSCs and studying the biology of reprogramming.

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Monolayer endoderm differentiation from human ESCs

Cited by 2 publications

References 0 publications

Generation of Self-Renewing Hepatoblasts From Human Embryonic Stem Cells by Chemical Approaches

Generation of Self-Renewing Hepatoblasts From Human Embryonic Stem Cells by Chemical Approaches

High-efficiency RNA-based reprogramming of human primary fibroblasts

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