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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。
OBJECTIVE: To explore the early hematopoietic reconstitution effect of co-transplantation of human placenta-derived mesenchymal stem cells (PMSCs) and cord blood mononuclear cells (MNCs) into SCID mice using intramedullary instillation of bone marrow (IBMI) technique. METHODS: Human PMSCs were obtained from human placenta by collagenase digestion, adherent culture and subculture. The surface markers were detected by flow cytometry (FCM) and induced to differentiate into osteoblasts and adipocytes. Osteogenic differentiation and identification of adipose tissue. The human PMSC and cord blood MNC were used as donors. The two kinds of cells were transplanted either in combination with IBMI or intravenous injection (IV) into the sublethal pretreated SCID mice were randomly divided into five groups: 50 in combination transplantation group (PMSC + cord blood MNC, IBMI), group B (cord blood MNC, IBMI), C transplantation group (PMSC via IBMI, Blood MNC via IV), saline control group D (after injection of saline only, IBMI), blank control group E (no irradiation only injection of saline, IBMI), 10 in each group. At 14 days after transplantation, bone marrow cells from the injected side and the contralateral tibial cavity of SCID mice in each group were collected. The levels of human CD34 + and CD45 + cells were detected by FCM. Results: PMSCs were successfully isolated and cultured from human placenta. FCM showed that the phenotype of PMSC was normal and successfully differentiated into osteoblasts and adipocytes. On the 14th day after SCID transplantation, only 4 mice survived , All other mice survived. The percentages of human CD34 + and CD45 + cells in the bone marrow of the contralateral and lateral tibia in group B were significantly lower than those in group A. Conclusion: Human PM-SC can improve the implantation rate of cord blood MNC and promote early hematopoietic reconstitution.