摘要
背景:目前,人脐血干细胞在脊髓损伤治疗中的研究较多,但人脐血干细胞的体外分化由许多因素所限制。蒙药广枣提取物对神经细胞有保护作用,但其作用机制不清楚。目的:观察蒙药广枣3种粗提取物促进人脐血干细胞体外分化的影响。方法:获取临床健康产妇的新鲜脐带血,制备人脐血干细胞悬液,将纯化的人脐血干细胞接种于40个培养皿中进行传代,根据蒙药广枣3种粗提取物分为:样品1组:乙醇提取,醋酸乙酯萃取,生药质量浓度为8.25g/mL;样品2组:乙醇提取,NKA树脂分离,体积分数为10%乙醇淋洗浓缩,生药质量浓度为1.72g/mL;样品3组:乙醇提取,NKA树脂分离,体积分数为70%乙醇淋洗浓缩,生药质量浓度为2.41g/mL;对照组:用80%的DMEM,含体积分数为20%的小牛血清培养。观察不同质量浓度蒙药广枣提取物对人脐血干细胞增殖的影响;24,72h后流式细胞仪测定S期的比例。结果与结论:样品3组人脐血干细胞的增殖作用不明显,10d时样品1组和样品2组明显高于样品3组和对照组(P<0.01)。样品1组和样品2组的人脐血干细胞在24,72h后处于S期的明显增多。提示生药质量浓度8.25,1.72g/mL蒙药广枣粗提取物能促进人脐血干细胞的体外增殖。
BACKGROUND:Human umbilical cord blood stem cells have been widely used in the study of spinal cord injury,but in vitro differentiation of human umbilical cord blood stem cells (hUCBSCs) has been limited by various factors.Mongalian medicine axillary choerospondias fruit extract has protective effects on neural cells,but the action mechanisms are unclear.OBJECTIVE:To observe promoting effects of 3 kinds of Mongalian medicine axillary choerospondias fruit extracts on in vitro differentiation of hUCBSCs.METHODS:Fresh umbilical cord blood was obtained from healthy puerperants to prepare hUCBSC suspension.The purified hUCBSCs were incubated in 40 petri dishes.The Mongalian medicine axillary choerospondias fruit extracts were divided into:sample 1 group:ethanol extraction,ethyl acetate extraction,crude drug mass concentration was 8.25 g/mL;sample 2 group:ethanol extraction,NKA resin isolation,10% ethanol eluting concentration,crude drug mass concentration was 1.72 g/mL;sample 3 group:ethanol extraction,NKA resin isolation,70% ethanol eluting concentration,crude drug mass concentration was 2.41 g/mL;control group:incubation of 80% DMEM containing 20% calf serum.Effects of various mass concentrations of Mongalian medicine axillary choerospondias fruit extract on hUCBSCs proliferation were observed.Proportion in S phase was measured using flow cytometry at 24 and 72 hours.RESULTS AND CONCLUSION:The proliferation of hUCBSCs was not significant in the sample 3 group.At day 10,the proliferation was significantly greater in the sample 1 and 2 groups compared with the sample 3 and control groups (P 0.01).The number of hUCBSCs was significantly increased at 24 and 72 hours in S phase in the sample 1 and 2 groups.Mongalian medicine axillary choerospondias fruit extract (crude drug mass concentration 8.25,1.72 g/mL) could promote in vitro proliferation of hUCBSCs.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2010年第6期1078-1081,共4页
Journal of Clinical Rehabilitative Tissue Engineering Research