Serum-free culturing of human mesenchymal stem cells with immobilized growth factors
Serum-free culturing of human mesenchymal stem cells with immobilized growth factors
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DOI：
10.1039/c6tb02867e
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发表时间：
2017-02-07
影响因子：
7
通讯作者：
Ito, Yoshihiro
中科院分区：
文献类型：
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作者：
Mao, Hongli;Kim, Seong Min;Ito, Yoshihiro
人骨髓间充质干细胞(HMSCs)的无血清体外扩增对基础研究和临床应用具有重要意义。已知，为hMSC培养开发特定的培养液需要一些生长因子。然而，当生长因子处于可扩散状态时，通常表现出稳定性差、循环半衰期短和细胞内化速度快的特点。克服这些问题的一个潜在方法是将生长因子固定在材料上。利用光反应明胶将碱性成纤维细胞生长因子、转化生长因子-β和血小板衍生生长因子三种不同类型的生长因子共固定在细胞培养皿表面，用于人骨髓间充质干细胞的无血清培养。结果表明，固定化生长因子对细胞增殖的支持作用与含血清培养基相类似。更重要的是，固定化生长因子显著提高了它们的热稳定性，有效地延长了它们在4℃和37℃下的保质期，而且固定化生长因子至少可以重复使用3次，而不会失去对细胞增殖的刺激作用。这种基于光反应明胶的生长因子固定化方法似乎是一种很有前途的无血清培养hMSC的方法。
In vitro expansion of human mesenchymal stem cells (hMSCs) using serum-free culture medium is important for basic research and clinical applications. It is known that some growth factors are required for developing a defined medium for hMSC culture. However, growth factors usually show poor stability, short circulating half-life and a rapid rate of cellular internalization when they are in a diffusible state. A potential way to overcome these problems is to immobilize growth factors on materials. Here, three different types of growth factors, basic fibroblast growth factor, transforming growth factor-beta, and platelet-derived growth factor, were co-immobilized on cell culture dish surfaces by photo-reactive gelatin and used for serum-free hMSC cultures. The results showed that the immobilized growth factors supported cell proliferation similarly to the serum-containing medium. More importantly, the immobilization of growth factors significantly improved their thermal stability and efficiently prolonged their shelf life at 4 degrees C and 37 degrees C. Furthermore, the immobilized growth factors could be reused at least three times without losing their stimulation effect on cell proliferation. This photo-reactive gelatin-based immobilization of growth factors appears to be a promising method for serum-free hMSC culturing.