Generation of induced pluripotent stem cell-derived beta-cells in blood amino acids-like medium.
Generation of induced pluripotent stem cell-derived beta-cells in blood amino acids-like medium.
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传统的细胞培养基不能准确地代表血浆中营养物质的可用性。它们通常含有超生理浓度的营养物质，例如葡萄糖、氨基酸等。这些高营养物质可以改变培养细胞的代谢并诱导不反映体内条件的代谢表型。我们证明营养物质的超生理水平会干扰内胚层分化。培养基配方的精制在体外干细胞衍生 β 细胞 (SC-β) 生成的成熟调节中具有潜在的应用。为了解决这些问题，我们建立了一个明确的培养系统，使用血液氨基酸样培养基 (BALM) 衍生 SC-β 细胞。在基于 BALM 的药物中，人诱导多能干细胞 (hiPSC) 可以有效分化为定形内胚层、胰腺祖细胞、内分泌祖细胞和 SC-β。分化的细胞在体外分泌 C 肽以响应高葡萄糖水平，并表达多种胰腺 β 细胞标记物。总之，生理水平的氨基酸足以产生功能性 SC-β 细胞。摘要：我们使用了类血液氨基酸培养基，发现它有助于有效地将人 iPSC 分化为定形内胚层，并生成功能性胰腺 β 细胞。
Traditional cell culture media do not accurately represent the availability of the nutrients in plasma. They usually contain a supraphysiological concentration of nutrients such as glucose, amino acids, etc. These high nutrients can alter the metabolism of cultured cells and induce metabolic phenotypes that do not reflect in vivo conditions. We demonstrate that the supraphysiological levels of nutrients interfere with endodermal differentiation. Refinement of media formulations has a potential application in maturity modulation of stem cell-derived β-cells (SC-β) generation in vitro. To address these issues, we established a defined culture system to derive SC-β-cells using a blood amino acids-like medium (BALM). Human induced pluripotent stem cells (hiPSCs) can be efficiently differentiated into the definitive endoderm, pancreatic progenitors, endocrine progenitors, and SC-β in BALM-based med. The differentiated cells secreted C-peptide in vitro in response to high glucose levels and expressed several pancreatic β-cell markers. In conclusion, amino acids at the physiological levels are sufficient for deriving functional SC-β cells. Summary: We used a blood amino acids-like medium and found that it is useful to efficiently differentiate human iPSCs into the definitive endoderm, and to generate functional pancreatic beta-cells.
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