当首次从原代培养的人乳牙牙髓细胞中获得iPSCs时,选择喂食器是很重要的。

I. Saitoh, E. Inada, Y. Iwase, H. Noguchi, Tomoya Murakami, Miki Soda, Naoko Kubota, H. Hasegawa, Eri Akasaka, Y. Matsumoto, K. Oka, Y. Yamasaki, H. Hayasaki, Masahiro Sato
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引用次数: 10

摘要

胚胎干细胞(ESCs)/诱导多能干细胞(iPSCs)的维持通常需要饲养细胞。小鼠胚胎成纤维细胞(MEFs)和STO小鼠基质细胞系是目前应用最广泛的饲养细胞。本研究的目的是确定哪些细胞适合从人乳牙牙髓细胞(HDDPCs)中构建多能干细胞。采用新型电穿孔法将HDDPCs原代培养物与含有人OCT3/4、SOX2/KLF4、LMYC/LIN28和pmaxGFP的三种质粒共转染,然后在ESC合格培养基中培养15 d。将新生菌落重新接种到丝裂霉素c处理的mef或STO细胞上。这些菌落被连续传代达26代。在此期间,评估集落形态,以确定细胞是否表现出esc样形态和碱性磷酸酶活性,以评估细胞重编程状态。维持在mef上的HDDPCs成功重编程为iPSCs,而维持在STO细胞上的HDDPCs则不能。一旦建立,iPSCs在STO细胞上维持而不丧失多能性。结果表明,mef细胞比STO细胞更适合培养iPSCs。馈线选择是高效生成iPSCs的关键因素。
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Choice of Feeders Is Important When First Establishing iPSCs Derived From Primarily Cultured Human Deciduous Tooth Dental Pulp Cells.
Feeder cells are generally required to maintain embryonic stem cells (ESCs)/induced pluripotent stem cells (iPSCs). Mouse embryonic fibroblasts (MEFs) isolated from fetuses and STO mouse stromal cell line are the most widely used feeder cells. The aim of this study was to determine which cells are suitable for establishing iPSCs from human deciduous tooth dental pulp cells (HDDPCs). Primary cultures of HDDPCs were cotransfected with three plasmids containing human OCT3/4, SOX2/KLF4, or LMYC/LIN28 and pmaxGFP by using a novel electroporation method, and then cultured in an ESC qualified medium for 15 days. Emerging colonies were reseeded onto mitomycin C-treated MEFs or STO cells. The colonies were serially passaged for up to 26 passages. During this period, colony morphology was assessed to determine whether cells exhibited ESC-like morphology and alkaline phosphatase activity to evaluate the state of cellular reprogramming. HDDPCs maintained on MEFs were successfully reprogrammed into iPSCs, whereas those maintained on STO cells were not. Once established, the iPSCs were maintained on STO cells without loss of pluripotency. Our results indicate that MEFs are better feeder cells than STO cells for establishing iPSCs. Feeder choice is a key factor enabling efficient generation of iPSCs.
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Cell medicine
Cell medicine MEDICINE, RESEARCH & EXPERIMENTAL-
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