1. A Synthetic Peptide-Acrylate Surface for Production of Insulin-Producing Cells from Human Embryonic Stem Cells
- Author
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Yao-Chen Yang, Shih-Han Hung, I-Ming Chu, Pei-Yi Lin, Li-Chuan Liao, Hsan-Jan Yen, Yi-Cheng Hsieh, Huai-En Lu, Shiaw-Min Hwang, and Maw-Sheng Lee
- Subjects
KOSR ,Surface Properties ,Cellular differentiation ,Cell Culture Techniques ,Biocompatible Materials ,Germ layer ,Biology ,Original Research Reports ,Humans ,Insulin ,Vitronectin ,Cells, Cultured ,Embryonic Stem Cells ,Matrigel ,Molecular Mimicry ,Cell Differentiation ,Cell Biology ,Hematology ,Molecular biology ,Embryonic stem cell ,Peptide Fragments ,Culture Media ,Drug Combinations ,Acrylates ,Cell culture ,Proteoglycans ,Collagen ,Laminin ,Subculture (biology) ,Stem cell ,Developmental Biology - Abstract
Human embryonic stem cells (hESCs), due to their self-renewal capacity and pluripotency, have become a potential source of transplantable β-cells for the treatment of diabetes. However, it is imperative that the derived cells fulfill the criteria for clinical treatment. In this study, we replaced common Matrigel with a synthetic peptide-acrylate surface (Synthemax) to expand undifferentiated hESCs and direct their differentiation in a defined and serum-free medium. We confirmed that the cells still expressed pluripotent markers, had the ability to differentiate into three germ layers, and maintained a normal karyotype after 10 passages of subculture. Next, we reported an efficient protocol for deriving nearly 86% definitive endoderm cells from hESCs under serum-free conditions. Moreover, we were able to obtain insulin-producing cells within 21 days following a simple three-step protocol. The results of immunocytochemical and quantitative gene expression analysis showed that the efficiency of induction was not significantly different between the Synthemax surface and the Matrigel-coated surface. Thus, we provided a totally defined condition from hESC culture to insulin-producing cell differentiation, and the derived cells could be a therapeutic resource for diabetic patients in the future.
- Published
- 2014