Effects of KnockOut Serum Replacement on Differentiation of Mouse-Induced Pluripotent Stem Cells into Odontoblasts
Effects of KnockOut Serum Replacement on Differentiation of Mouse-Induced Pluripotent Stem Cells into Odontoblasts
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DOI：
10.2209/tdcpublication.2021-0042
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发表时间：
2022-01-01
影响因子：
0.5
通讯作者：
Azuma, Toshifumi
中科院分区：
文献类型：
--
作者：
Furukawa, Yuki;Odashima, Ayano;Azuma, Toshifumi
血清是体外细胞培养过程中丰富营养的来源，促进细胞粘附、生长和分化。然而，当培养用于移植的干细胞时，必须记住，这样的培养基可能含有对拟接受者潜在有害的物质，甚至可能诱导细胞损伤。本研究的目的是确定是否敲除血清替代品（KSR），化学成分确定的培养基补充剂，在体外诱导多能干细胞分化成成牙本质细胞的增强。颅神经嵴细胞是成牙本质细胞的前体细胞，由小鼠诱导的多能干细胞产生。然后将它们培养在含有成纤维细胞生长因子8的无血清Dulbecco改良的Eagle/F12培养基中，有或没有KSR。KSR培养的细胞增殖旺盛，呈梭形，与周围细胞连接。通过qRT-PCR测量，KnockOut血清替代物还促进了成牙本质细胞标记物的表达，并通过免疫染色评估增加了牙本质唾液蛋白。这些结果证实了小鼠诱导的多能干细胞在无血清条件下分化为成牙本质细胞，并且KSR增强了这一过程的效率。
Serum serves as a source of rich nutrients during in vitro cell culture, facilitating cell adhesion, growth, and differentiation. When culturing stem cells for transplantation, however, it must be remembered that such culture medium may contain substances potentially harmful to the proposed recipient and may even induce cellular damage. The purpose of this study was to determine whether KnockOut Serum Replacement (KSR), a chemically defined medium supplement, enhanced in vitro differentiation of induced pluripotent stem cells into odontoblasts. Cranial neural crest cells, precursors of odonto-blasts, were generated from mouse-induced pluripotent stem cells. They were then cul-tured in serum-free Dulbecco's modified Eagle's/F12 medium containing fibroblast growth factor 8 with or without KSR. The cells cultured with KSR showed strong prolifera-tion, acquired a spindle-like morphology, and connected with the surrounding cells. KnockOut Serum Replacement also boosted expression of odontoblast markers as mea-sured by qRT-PCR, and increased dentin sialoprotein as assessed by immunostaining. These results confirmed that mouse-induced pluripotent stem cells differentiated into odontoblasts under serum-free conditions, and that KSR enhanced the efficiency of this process.