Peh Gary S L, Lee Man-Xin, Wu Fei-Yi, Toh Kah-Peng, Balehosur Deepashree, Mehta Jodhbir S
Singapore Eye Research Institute, Singapore 168751.
Int J Biomater. 2012;2012:601302. doi: 10.1155/2012/601302. Epub 2012 Jan 12.
The culture of human corneal endothelial cells (CECs) is critical for the development of suitable graft alternative on biodegradable material, specifically for endothelial keratoplasty, which can potentially alleviate the global shortage of transplant-grade donor corneas available. However, the propagation of slow proliferative CECs in vitro can be hindered by rapid growing stromal corneal fibroblasts (CSFs) that may be coisolated in some cases. The purpose of this study was to evaluate a strategy using magnetic cell separation (MACS) technique to deplete the contaminating CSFs from CEC cultures using antifibroblast magnetic microbeads. Separated "labeled" and "flow-through" cell fractions were collected separately, cultured, and morphologically assessed. Cells from the "flow-through" fraction displayed compact polygonal morphology and expressed Na(+)/K(+)ATPase indicative of corneal endothelial cells, whilst cells from the "labeled" fraction were mostly elongated and fibroblastic. A separation efficacy of 96.88% was observed. Hence, MACS technique can be useful in the depletion of contaminating CSFs from within a culture of CECs.
人角膜内皮细胞（CECs）的培养对于在可生物降解材料上开发合适的移植替代物至关重要，特别是对于内皮角膜移植术，这有可能缓解全球可用的移植级供体角膜短缺的问题。然而，在某些情况下可能会共同分离出快速生长的角膜基质成纤维细胞（CSFs），从而阻碍体外缓慢增殖的CECs的增殖。本研究的目的是评估一种使用磁性细胞分离（MACS）技术的策略，该技术使用抗成纤维细胞磁性微珠从CEC培养物中去除污染的CSFs。分别收集分离出的“标记”和“流通”细胞组分，进行培养并进行形态学评估。“流通”组分中的细胞呈现紧密的多边形形态，并表达指示角膜内皮细胞的Na(+)/K(+)ATPase，而“标记”组分中的细胞大多呈细长形且为成纤维细胞样。观察到的分离效率为96.88%。因此，MACS技术可用于从CEC培养物中去除污染的CSFs。
