论文部分内容阅读
目的研究体外分离培养大鼠外周血基质干细胞(PBMSCs)并诱导分化为施万细胞的潜能。方法从SD大鼠取血分离培养PBMSCs,采用流式细胞术和免疫细胞化学法对其细胞表面抗原进行检测与鉴定,并用免疫细胞化学法检测BrdU作用4h后BrdU的阳性率。PBMSCs经β-巯基乙醇、全反式维甲酸及复合条件培养基等三个步骤向施万细胞定向诱导后,用免疫细胞化学法检测S-100和P75的表达。结果流式细胞术显示培养获得的PBMSCs中CD11b、CD29、CD45、CD49d、CD90及CD106的阳性率分别为19.97%、99.96%、46.62%、5.46%、71.22%和10.76%。免疫细胞化学法显示PBMSCs呈CD34阴性,而BrdU阳性率为(34.1±4.3)%。PBMSCs经定向诱导后S-100和P75的阳性率分别是(75.2±4.1)%和(78.9±4.6)%。结论从外周血分离获得的干细胞符合基质干细胞的特性,这些细胞在特定的条件下可诱导分化成为施万细胞。
Objective To study the potential of isolated and cultured rat peripheral blood stromal stem cells (PBMSCs) in vitro and to differentiate into Schwann cells. Methods PBMSCs were isolated and cultured from SD rats. The cell surface antigens were detected by flow cytometry and immunocytochemistry. The positive rate of BrdU after 4 hours was detected by immunocytochemistry. PBMCs were induced into Schwann cells by three steps, such as β-mercaptoethanol, all-trans retinoic acid and compound conditioned media. The expression of S-100 and P75 was detected by immunocytochemistry. Results Flow cytometry showed that the positive rates of CD11b, CD29, CD45, CD49d, CD90 and CD106 in PBMSCs were 19.97%, 99.96%, 46.62%, 5.46%, 71.22% and 10.76% respectively. Immunocytochemistry showed that PBMSCs were CD34 negative, while the positive rate of BrdU was (34.1 ± 4.3)%. The positive rates of S-100 and P75 induced by PBMCs were (75.2 ± 4.1)% and (78.9 ± 4.6)%, respectively. Conclusion The stem cells isolated from peripheral blood are in accordance with the characteristics of stromal stem cells. Under certain conditions, these cells can be induced to differentiate into Schwann cells.