CD200 facilitates the isolation of corneal epithelial cells derived from human pluripotent stem cells.
CD200 facilitates the isolation of corneal epithelial cells derived from human pluripotent stem cells.
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DOI：
10.1038/s41598-018-34845-2
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发表时间：
2018-11-08
影响因子：
4.6
通讯作者：
Nishida K
中科院分区：
文献类型：
--
作者：
Hayashi R;Ishikawa Y;Katayama T;Quantock AJ;Nishida K
从人诱导多能干细胞（iPSC）体外诱导角膜上皮细胞（CEC）代表了获得CE干细胞/祖细胞用于手术重建患病或受伤眼表面的新策略。这种策略的临床前景是相当可观的，但如果要实现这些方法的潜力，则需要开发用于纯化iPSC衍生的CE谱系细胞的稳健方法，以避免与可能带来不必要的副作用风险的其他细胞污染，例如肿瘤发生。在此进行的实验显示，在CEC分离期间，与使用TRA-1-60（CEC的常规阴性标记物）可以实现的情况相比，使用细胞分选仪的CD 200阴性选择显著减少了细胞群被各种非CEC污染。此外，CD 200阴性分选不影响CEC的产量，也不影响其干/祖细胞的产量。使用CD 200阴性分选获得的CEC片的单细胞基因表达分析显示，所有分析的细胞都是CE谱系细胞，表达PAX 6、Δ-Np 63和E-钙粘蛋白。另一方面，非CEC表达非CEC基因，如FGFR 1和RPE 65。因此，CD 200代表用于纯化诱导的CE谱系细胞的稳健阴性标志物，其由未分化的iPSC和非CEC（包括iPSC衍生的神经和视网膜细胞）表达。
The in vitro induction of corneal epithelial cells (CECs) from human induced pluripotent stem cells (iPSCs) represents a new strategy for obtaining CE stem/progenitor cells for the surgical reconstruction of a diseased or injured ocular surface. The clinical promise of this strategy is considerable, but if the approaches’ potential is to be realised, robust methods for the purification of iPSC-derived CE lineage cells need to be developed to avoid contamination with other cells that may carry the risk of unwanted side effects, such as tumorigenesis. Experiments conducted here revealed that during CEC isolation, CD200-negative selection using a cell sorter considerably reduced the contamination of the cell population with various non-CECs compared with what could be achieved using TRA-1-60, a conventional negative marker for CECs. Furthermore, CD200-negative sorting did not affect the yield of CECs nor that of their stem/progenitor cells. Single-cell gene expression analysis for CEC sheets obtained using CD200-negative sorting showed that all analysed cells were CE-lineage cells, expressing PAX6, delta-N p63, and E-cadherin. Non-CECs, on the other hand, expressed non-CEC genes such as FGFR1 and RPE65. CD200, thus, represents a robust negative marker for purification of induced CE lineage cells, which is expressed by undifferentiated iPSCs and non-CECs, including iPSC-derived neural and retinal cells.
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影响因子：
5.9
作者：
Doi, Daisuke;Samata, Bumpei;Katsukawa, Mitsuko;Kikuchi, Tetsuhiro;Morizane, Asuka;Ono, Yuichi;Sekiguchi, Kiyotoshi;Nakagawa, Masato;Parmar, Malin;Takahashi, Jun
通讯作者：
Takahashi, Jun
影响因子：
5.6
作者：
Büttner, C;Skupin, A;Rieber, EP
通讯作者：
Rieber, EP
影响因子：
16.2
作者：
Pirvola, U;Ylikoski, J;Partanen, J
通讯作者：
Partanen, J
影响因子：
16.6
作者：
通讯作者：
--
影响因子：
3
作者：
Chen, Szu-Yu;Hayashida, Yasutaka;Tseng, Scheffer C. G.
通讯作者：
Tseng, Scheffer C. G.