A 3D culture system enhances the ability of human bone marrow stromal cells to support the growth of limbal stem/progenitor cells
A 3D culture system enhances the ability of human bone marrow stromal cells to support the growth of limbal stem/progenitor cells
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DOI：
10.1016/j.scr.2016.02.018
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发表时间：
2016-03-01
影响因子：
1.2
通讯作者：
Deng, Sophie X.
中科院分区：
文献类型：
--
作者：
Gonzalez, Sheyla;Mei, Hua;Deng, Sophie X.
在单层小鼠 3T3 饲养细胞上培养角膜缘上皮祖细胞/干细胞 (LSC) 的标准方法在临床应用中存在交叉污染的风险。人类饲养细胞已被用来消除这种风险；然而，单层无外源培养物的效率似乎低于使用 3T3 细胞的标准方法。我们研究了骨髓基质细胞 (BMSC)（也称为骨髓源性间充质干细胞）是否可以作为 LSC 在 3 维 (3D) 系统中扩增的饲养细胞。单层 3T3 上的原代单个人类 LSC 作为对照。当在 BMSC 上培养单个 LSC 时，观察到生长非常差。当 LSC 簇在 BMSC 单层 (CC-BM)、3D 培养系统 (3D CC-BM) 和纤维蛋白 3D 系统 (纤维蛋白 3D CC-BM) 上培养时，3D CC-BM 方法支持更大的 LSC 扩增。 3D CC-BM 系统产生的细胞生长速率比对照高 2.5 倍 (p < 0.05)。 K14(+)和p63α(bright)细胞的比例与对照组相当(p>0.05)，而K12(+)细胞的比例较低(p<0.05)。这些结果表明 BMSC 可以有效支持 3D 培养中 LSC 群体的扩张。 (C) 2016 年作者。由 Elsevier B.V. 出版
The standard method of cultivating limbal epithelial progenitor/stem cells (LSCs) on a monolayer of mouse 3T3 feeder cells possesses the risk of cross-contamination in clinical applications. Human feeder cells have been used to eliminate this risk; however, efficiency from xenobiotic-free cultures on a monolayer appears to be lower than in the standard method using 3T3 cells. We investigated whether bone marrow stromal cells (BMSCs), also known as bone marrow-derived mesenchymal stem cells, could serve as feeder cells for the expansion of LSCs in the 3-dimensional (3D) system. Primary single human LSCs on a monolayer of 3T3s served as the control. Very poor growth was observed when single LSCs were cultured on BMSCs. When LSC clusters were cultured on a BMSC monolayer (CC-BM), 3D culture system (3D CC-BM) and fibrin 3D system (fibrin 3D CC-BM), the 3D CC-BM method supported a greater LSC expansion. The 3D CC-BM system produced a 2.5-fold higher cell growth rate than the control (p < 0.05). The proportion of K14(+) and p63 alpha(bright) cells was comparable to those in the control (p > 0.05), whereas the proportion of K12(+) cells was lower (p < 0.05). These results indicate that BMSCs can efficiently support the expansion of the LSC population in the 3D culture. (C) 2016 The Authors. Published by Elsevier B.V.