摘要
目的优化脐血造血干细胞/祖细胞的分离方法。方法脐血按体积比1∶5与含有树脂海绵筛(筛孔直径约12μm)的6%羟乙基淀粉混合,静置60min,提取界面有核细胞。倒置显微镜下计数有核细胞数,集落培养记数CFU-GM数,流式细胞仪检测CD34+细胞数,并与按照标准的羟乙基淀粉沉淀法和淋巴细胞分离液密度梯度离心法分离的脐血相对照。结果采用本实验方法分离脐血获得的有核细胞数、CFU-GM数和CD34+细胞数分别为(96.82±25.78)×105/ml,(66.70±12.47)×105,(1.33±0.57)×105/ml,明显高于由羟乙基淀粉沉淀法(70.72±20.93)×105/ml,(28.79±10.25)×105,(1.01±0.36)×105/ml和淋巴细胞分离液密度梯度离心法(25.33±7.27)×105/ml,(20.13±7.55)×105,(0.34±0.01)×105/ml分离的脐血(P<0.05)。结论本实验方法可由脐血中分离出大量的造血干/祖细胞,是一种分离脐血造血干/祖细胞的理想方法。
Objective To optimize the protocols for isolating hematopoietic stem/progenitor cells （HSPC） from umbilical cord blood （UCB）. Methods Nucleated cells （NC） derived from UCB were separated by hydroxyethyl starch （6% HES） which mixed with resin sponge （pore 12urn）. The number of NC,CFU- GM and CD34^＋ cells were detected by enumering, colony cuture and flow cytometery, and compered with that from UCB seperated by standard hydroxyethyl starch （6% HES） sedimentation or density gradient centrifugation （ Ficoll - Hypaque, 1. 007g/L）. Results The number of NC, CFU - GM and CD34^＋ cells isolated by our medthod were （96.82 ± 25.78） × 10^5/ml, （66.70 ± 12.47） × 10^5 and （ 1.33 ± 0.57） × 10^5/ml respectively, which were higher than that from UCB isolated by standard hydroxyethy starch sedimentation （ 70.72 ± 20.93 ） × 10^5/ml; （28.79 ± 10.25 ） × 10^5, （ 1.01 ± 0.36） × 10^5/ml or density gradient centrifugation （ 25.33 ± 7.27） × 10^5/ml, （20.13 ±7.55） × 10^5,（0.34 ±0.01） ×10^5/ml （ P 〈0.05）. Conclusion It is concluded that the protocols established here can acquire HSPC with high abundance, enrichment and viability and may be a optimal method for isolating hematopoietic stern/progenitor cells from umbilical cord blood.
出处
《医药论坛杂志》
2007年第5期52-54,共3页
Journal of Medical Forum
关键词
脐血
造血干细胞
细胞分离
Umbilical cord blood
Hematopoietic stem cell
Cell separation
