摘要
目的 建立获取高纯度人骨髓来源的多能成体祖细胞(MAPCs)的方法。方法 取志愿者适量骨髓后采用梯度密度离心法分离获取单个核细胞,对单个核细胞行贴壁培养后,将获取的骨髓贴壁细胞通过CD45、Gly-A免疫做磁珠负分选,流式细胞仪鉴定分选后提高细胞纯度。结果 (1)通过MAPCs分选,半均每1×106／mL骨髓贴壁细胞可分选出约(5.1±2.8)×104／mL数量级的MAPCs,分选前后细胞活力分别为96.7％±1.7％和96.0％±2.4％,无明显差异;(2)流式细胞仪分析获取的CD45-、Gly-A-细胞纯度大于98％。结论 CD45、Gly-A免疫微磁珠负分选可从骨髓单个核细胞中中分选出高纯度的MAPCs。
Objective To design a method of obstaining the highly-purified multipotent adult progenitor cells (MAPCs) from human bone marrow in vitro. Methods The mononuclear cells were isolated from the proper volume of bone marrow from volunteers by density gradient centrifugation, and then were cultivated by adherent culture to collect the bone marrow adherent cells. The adherent cells were selected and separated by use of CD45 and Gly-A immunomagnetic beads. Analyze the component of the CD45-, Gly-A- cells by flow cytometer. Results (1)Per 1×106/mL bone marrow adherent monocytes, (5.1± 2.8)×104/mL MAPCs could be separated by magnetic activated cell sorting (MACS). The activity of cells before and after separation was 96.7%±1.7% and 96.0%±2.4% respectively. (2)The purity of the CD45-、Gly-A cells separated from bone marrow adherent cells was more than 98% by flow cytometers. Conclusion The MAPCs derived from bone marrow can be purified highly by use of CD45 and Gly-A immunomagnetic beads.
出处
《中华神经医学杂志》
CAS
CSCD
2004年第6期422-424,共3页
Chinese Journal of Neuromedicine
基金
广东省科技计划项目(2002A3020206)
