State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, GuangZhou 510060, China.
Sci Rep. 2013;3:1933. doi: 10.1038/srep01933.
We isolated human epidermis-derived mesenchymal stem cell-like pluripotent cells (hEMSCPCs) and demonstrate efficient harvesting, maintenance in vitro for at least 30 passages, reprogramming into multiple phenotypes in vivo, and integration into adult host tissues after injection into the mouse blastocyst to create chimeras. Cell phenotype was examined by karyotyping, immunostaining, immunofluorescence, and flow cytometry. A nested PCR protocol using primers specific for human SRY genes was designed to detect hEMSCPC-derived cells in female chimeric mice. FISH was used to validate the results of nested PCR. Results indicated that hEMSCPCs were derived from epidermis but were distinct from epidermal cells; they resembled mesenchymal stem cells (MSCs) morphologically and expressed the main markers of MSCs. About half of all female offspring of mice implanted with embryos injected with hEMSCPCs at the blastocyst stage harbored the human Y chromosome and tissue-specific human protein, thereby demonstrating the transdifferentiation of hEMSCPCs.
我们分离了人表皮来源的间充质干细胞样多能细胞（hEMSCPCs），并证明其能够高效收获和体外维持至少 30 代，在体内重编程为多种表型，并在注射到小鼠囊胚中创建嵌合体后整合到成年宿主组织中。通过核型分析、免疫染色、免疫荧光和流式细胞术检查细胞表型。设计了一种使用针对人 SRY 基因的特异性引物的嵌套 PCR 方案，以检测雌性嵌合小鼠中的 hEMSCPC 衍生细胞。FISH 用于验证嵌套 PCR 的结果。结果表明，hEMSCPCs 来源于表皮，但与表皮细胞不同；它们在形态上类似于间充质干细胞（MSCs），并表达 MSCs 的主要标记物。大约一半在囊胚阶段注射 hEMSCPCs 的小鼠的雌性后代携带人类 Y 染色体和组织特异性人类蛋白，从而证明了 hEMSCPCs 的转分化。
