中国组织工程研究与临床康复2011，Vol.15Issue(45)：8447-8450,4.DOI:10.3969/j.issn.1673-8225.2011.45.019
两步法分离人脐血单个核细胞的最佳条件
An exploration of the optimal conditions for isolating umbilical cord blood mononuclear cells by using two-step methods
郭继强 1刘爱兵 1沈丹1
作者信息
- 1. 武警总医院医学实验中心,北京市,100039
- 折叠
摘要
Abstract
BACKGROUND: It has not been deeply reported howto obtain high-purity and high-vigor stem cells as well as the standardizedoperation plan has not been seen, yet.OBJECTIVE: To explore the best condition for isolating human umbilical cord blood mononuclear cells.METHODS: Umbilical cord blood red blood cells precipitated by hydroxyethyl starch were observed in different time points (20,30, 40, 50, 60, 70 minutes). The human umbilical cord blood mononuclear cells were separated in human lymphocyte separatingmedium (LSM) by using centrifugation method at 800, 700, 600, 500, 400 g/min for 30, 25, and 20 minutes.RESULTS AND CONCLUSION: The 6% hydroxyethyl starch precipitation for umbilical cord blood red blood cells was best within60 minutes. Using the human LSM of (1.077 0+0.000 1) g/mL at 4 ℃, centrifugation at 700 g/min for 30 minutes, and rinsing for 3times the best conditions to harvest human umbilical cord blood mononuclear cells. Under the above-mentioned conditions, notonly higher umbilical cord blood mononuclear cells rate and higher mononuclear cells vigor were obtained, but fewer red bloodcells and other cell fragments were seen. The best condition for isolating the human umbilical cord blood mononuclear cells usingtwo steps methods (6% hydroxyethyl starch precipitation and LSM) at the optimal time can improve the recovery rate of humanumbilical cord blood stem cells.关键词
人脐血/单个核细胞/密度梯度离心法/羟乙基淀粉/两步法分类
基础医学引用本文复制引用
郭继强,刘爱兵,沈丹..两步法分离人脐血单个核细胞的最佳条件[J].中国组织工程研究与临床康复,2011,15(45):8447-8450,4.
