摘要
目的 :为了建立免疫磁珠法体外分离提纯骨髓造血干细胞的方法。方法 :采用免疫磁珠阴性选择法分选 CBA(H - 2 K)小鼠骨髓细胞的 CD34+ CD38- Scal+ 细胞群 ,台盼蓝染色观察活力 ,流式细胞仪检测荧光标记的 CD34+ CD38- Scal+ 表达阳性率 ,骨髓造血干细胞 (HSCs)在体外克隆形成的分析 ,HSCs在体内造血功能重建的分析。结果 :分选后 CD34+ CD38- Scal+ 表达阳性细胞纯度达 (86 .5± 3.2 ) % ,细胞活力为 (95 .8± 2 .2 ) %。结论 :免疫磁珠阴性选择法分选系统是一种有效的骨髓造血干细胞分离提纯方法 ,省时、简便、纯度高并对细胞的生物活性无明显影响 。
Objective:To isolate and purify murine hematopoietic stem cell using the immunomagnetic microbeads.Methods:CBA(H-2 K)mice HSC from bone marrow cells were isolated by negative selection using magnetically labeled mAbs. The HSCs purification and their viability were determined by flow cytomety and light microscopy, respectively.The assessment of hematopoiesis was evaluated through forming colonies in M3434 semisolid and HSC being transplanted into myeloablated syngenetic mice,respectively.Result:CD34 +CD38 -Scal + HSCs and their viability were (86.5±3.2)%,(95.8±2.2)%, respectively.BFU-E, CFU-GM and CFU-GMM were formed in M3434 semisolid. All the mice transplanted CD34 +CD38 -Scal + HSCs after lethal irradiation survived, and all the mice in control groups died two weeks later.Conclusion:It is effetive to isolated HSCs from murine bone marrow cells by negative selection using magnetically labeled mAbs, and the purity and viability of the cells are high.
出处
《广西医科大学学报》
CAS
2004年第4期478-480,共3页
Journal of Guangxi Medical University
关键词
免疫磁珠法
分离
提纯
骨髓造血干细胞
immunomagnetic beads
hematopoietic stem cell
isolation and purification
