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免疫磁珠法及羟乙基淀粉法分离脐血单个核细胞的比较
李敏, 范淑英, 孙光辉, 徐佩茹, 阿尔孜古丽·吐尔逊
中国当代儿科杂志 ›› 2009, Vol. 11 ›› Issue (9) : 757-760.
PDF(1383 KB)
PDF(1383 KB)
免疫磁珠法及羟乙基淀粉法分离脐血单个核细胞的比较
Comparison of immunomagnetic beads and hespan precipitation for isolation of mononuclear cells from umbilical cord blood
目的：该实验比较了CD133免疫磁珠法及羟乙基淀粉法分离脐血单个核细胞（MNCs）的特点，探讨一种相对较好的MNCs分离方法。方法：取2007年10月至2008年3月新疆医科大学第一附属医院足月妊娠健康产妇的脐血15份，每份脐血分别用羟乙基淀粉沉淀法、CD133免疫磁珠法处理。分离后计数并在倒置显微镜下观察原代细胞的生长情况及其形态特征，原代第30天通过流式细胞仪检测CD34阳性率。结果：羟乙基淀粉沉淀法、CD133免疫磁珠组得到的MNCs数量分别为（15.23±4.30）×106/mL，（0.066±0.027）×106/mL（P＜0.05）。羟乙基淀粉沉淀法所得细胞大多悬浮生长，传代后生长缓慢；CD133免疫磁珠法的细胞生长状态良好，其CD34阳性率分别为10.1％、0.5％。结论：羟乙基淀粉沉淀法是一种高效的脐血细胞分离方法，但细胞生长状态欠佳；而 CD133免疫磁珠法所获细胞纯度较高，可根据不同要求选用相应分离法。［中国当代儿科杂志，2009，11（9）：757-760］
OBJECTIVE: To compare the characteristics of immunomagnetic beads and hespan precipitation for isolation of mononuclear cells (MNCs) from umbilical cord blood and try to find a better isolation method for MNCs. METHODS: Fifteen umbilical cord blood samples from healthy parturiens were collected between December 2007 and March 2008. MNCs were isolated using hespan precipitation and CD133 immunomagnetic beads, respectively. MNCs were identified using the surface marker CD34 by flow cytometry on the 30th of primary culture. Growth conditions and morphologic changes of primary cells were observed by an inverted microscope. RESULTS: The number of MNCs from umbilical cord blood isolated by hespan precipitation (15.23±4.30×106/mL) was significantly greater than that by CD133 immunomagnetic beads (0.066±0.027×106/mL) (P<0.05). The MNCs isolated by hespan precipitation suspended at the culture medium and their growth was slow after passage. The growth of MNCs isolated by CD133 immunomagnetic beads was kept in a good condition. The CD34 positive rate of MNCs isolated by hespan precipitation and immunomagnetic beads was 10.1% and 0.5%, respectively. CONCLUSIONS: The hespan precipitation is an effective method for MNCs isolation from human umbilical cord blood, but with a cell growth condition below the mark. The MNCs isolated by CD133 immunomagnetic beads are in a high purity quotient.［Chin J Contemp Pediatr, 2009, 11 (9):757-760］
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