摘要
目的:利用骨髓腔内注射(IBMI)技术初步探讨人胎盘源间充质干细胞(PMSC)联合脐血单个核细胞(MNC)共移植入SCID鼠体内后的早期造血重建效应。方法:将人胎盘组织经胶原酶消化、贴壁和传代培养获得人PMSC,运用流式细胞仪术(FCM)检测其表面标志,并将其诱导分化为成骨细胞和脂肪细胞,继而对其进行成骨分化、脂肪分化鉴定;以人PMSC和脐血MNC为供体,通过IBMI方法或结合静脉注射(IV)方法,将两种细胞联合或单独移植入经亚致死量辐照预处理的SCID受鼠体内,50只受鼠随机分为5组:联合移植A组(PMSC+脐血MNC,IBMI)、单独移植B组(脐血MNC,IBMI)、联合移植C组(PMSC经IBMI,脐血MNC经IV)、生理盐水对照D组(辐照后仅注射生理盐水,IBMI)、空白对照E组(不辐照仅注射生理盐水,IBMI),每组10只。在移植后14 d取各组SCID小鼠注射侧和对侧胫骨腔内骨髓细胞,并用FCM检测分析人CD34+、CD45+细胞的植入水平。结果:从人胎盘组织中成功分离和培养PMSC,FCM检测其表型正常,并成功向成骨和成脂肪细胞诱导分化;SCID小鼠移植后14 d,照射对照组小鼠仅剩4只存活,其他组小鼠全部存活,B组注射侧及对侧胫骨骨髓腔内的人CD34+、CD45+细胞的百分比均明显低于A组小鼠的注射侧和对侧。结论:人PM-SC可以提高脐血MNC的植入率,促进早期造血重建。
AIM： To evaluate the early hematopoietic reconstitution of implanted SCID mice which were transplanted with human umbilical cord blood-derived mononuclear cells（UCB-MNC） and placenta-derived mesenchymal stem cells（PMSC） by intro-bone marrow injection（IBMI）.METHODS： The placenta tissues were digested by collagenase IV and cultured with low-glucose DMEM supplemented with b-FGF.The adherent cells were collected and passaged.The phenotypes of the cultured cells were detected by flow cytometry.The osteogenic differentiation and adipogenic differentiation were induced and detected.SCID recipient mice conditioned with sublethal dose irradiation were transplanted with human UCB-MNC and PMSC by IBMI or intravenous injection（IV）.Fifty recipient mice were divided into five groups at random： cotransplantation group A（PMSC＋UCB-MNC by IBMI）,single transplantation group B（UCB-MNC by IBMI）,cotransplantation group C（PMSC by IBMI,UCB-MNC by IV）,normal saline control group D（normal saline by IBMI）,normal control group E（normal saline by IBMI）.There were ten recipient mice in every group.On day 14,the bone marrow cells of every recipient mouse were flushed out from the injected tibias and contralateral tibias,respectively.The percentage of human CD34＋ and CD45＋ hematopoietic cells was analyzed by flow cytometry.RESULTS： PMSC were isolated and expanded from human placenta,which were successfully induced to osteogenic differentiation and adipogenic differentiation.FACS analyses showed that the phenotypes of PMSC were normal.On day 14 after transplantation in SCID mice,the percentages of human CD34＋ and CD45＋ hematopoietic cells in the tibias of group B were both significantly lower than them in the injected tibias and contralateral tibias of group A.CONCLUSION： Human PMSC could enhance the early engraftment of UCB-MNC in SCID mice.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2010年第11期1097-1100,共4页
Chinese Journal of Cellular and Molecular Immunology
基金
国家自然科学基金资助项目(30400395)
关键词
胎盘源间充质干细胞
造血重建
骨髓腔内注射
脐血单个核细胞
placenta-derived mesenchymal stem cell
hematopoietic reconstitution
intra-bone marrow injection
umbilical cord blood-derived mononuclear cell
