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通过含氮等离子体提高改性聚苯乙烯对脐带间充质干细胞的黏附与增殖

Improving the attachment and proliferation of umbilical cord mesenchymal stem cells on modified polystyrene by nitrogen-containing plasma.

机构信息

The Graduate School, Chiang Mai University, 239 Huaykaew Rd., Muang, 50200, Thailand.

出版信息

Cytotechnology. 2013 Jan;65(1):119-34. doi: 10.1007/s10616-012-9467-9. Epub 2012 Jul 4.

Abstract

Wharton's jelly mesenchymal stem cells (WJMSCs) are important alternative source of pluripotent cells for several therapeutic purposes. Understanding of adhesion properties of such cells is necessary to regulate the attachment, growth and proliferation on targeted culture surfaces. BCP-K1, a line of WJMSCs, and polystyrene (PS) culture dishes were used as membrane samples. A 13.56 MHz inductively coupled discharge plasma reactor with a mixture of N-containing gas and noble gas was used. This was expected to introduce the more hydrophilic groups on PS surface and enhance the cell adhesion. The plasma-treated PS dishes with the mixed gas of N(2) + He at 50 W and NH(3) + He at 100 W were reactive towards BCP-K1. Cellular adhesion and proliferation was significantly twice as efficient on the treated surfaces than on PS dishes. BCP-K1 also secreted more focal adhesion kinase to adhere and proliferate when cultured on N(2)-treated PS dishes than on the NH(3)-treated PS dishes. Stable stemness markers were detected, including CD105, CD9 and SSEA-4, expressed on BCP-K1 growing on the modified PS dish surfaces, during 7 days of culturing. The presence of -NH(2) groups on the PS dish surface were revealed by X-ray photoelectron spectroscopy and Fourier transform infrared spectroscopy. A large amount of oxygen- and nitrogen-containing functional groups, up to 9.0 %, were introduced by NH(3) plasma and N(2) plasma. The functional groups introduced on to the PS surfaces were clearly the key factors which enhanced WJMSCs attachment and stemness stability.

摘要

Wharton 胶间充质干细胞(WJMSCs)是多种治疗用途中多能细胞的重要替代来源。为了调节细胞在目标培养表面的附着、生长和增殖,了解这些细胞的粘附特性是必要的。BCP-K1 是一种 WJMSC 系,聚苯乙烯(PS)培养皿被用作膜样品。使用了一种混合含氮气体和稀有气体的 13.56MHz 感应耦合放电等离子体反应器。这有望在 PS 表面引入更多亲水性基团,增强细胞粘附性。在 50W 的 N2+He 和 100W 的 NH3+He 混合气体下处理的 PS 盘对 BCP-K1 具有反应性。处理过的表面上细胞粘附和增殖的效率比 PS 盘高两倍。当在 N2 处理过的 PS 盘上培养时,BCP-K1 分泌更多的粘着斑激酶来粘附和增殖,而不是在 NH3 处理过的 PS 盘上。在培养的 7 天内,在经过修饰的 PS 盘表面上生长的 BCP-K1 上检测到稳定的干细胞标志物,包括 CD105、CD9 和 SSEA-4。X 射线光电子能谱和傅里叶变换红外光谱表明 PS 盘表面存在 -NH2 基团。NH3 等离子体和 N2 等离子体引入了高达 9.0%的大量含氧和含氮官能团。引入 PS 表面的官能团显然是增强 WJMSCs 附着和干细胞稳定性的关键因素。

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