不同供者脐带间充质干细胞生物学特性的比较研究

发布时间：2019-04-24 03:58

【摘要】：脐带间充质干细胞(UC-MSCs)具有自我更新、增殖和分化的潜能，在造血恢复、修复损伤或病变的组织器官、免疫抑制治疗等方面具有重要临床应用前景。为了获得稳定质量的UC-MSCs，研究不同供者UC-MSCs生物学特性的差异是十分必要的。为此，我们对脐带的存放时间和UC-MSCs的培养进行研究，通过观察不同供者来源的UC-MSCs的细胞形态、增殖能力、多向分化能力、表面分子标志物和细胞因子的表达量，比较其生物学特性的差异。用UC-MSCs培养上清，比较不同供者来源UC-MSCs促造血因子的分泌能力。采集后的新鲜脐带存放时间不宜超过48h。脐带存放时间48~72h时，分离UC-MSCs的成功率下降为80%，细胞数减少为3.83×10~4/cm、原代培养时间延长为10.8d。DMEM/F12是UC-MSCs的最适培养基，原代培养时间为8.24d，获得的细胞数达3.84×10~6。20~31岁的不同供者的UC-MSCs体外培养均呈纺锤形、贴壁生长，在诱导培养基中均能分化为脂肪细胞和成骨细胞，表达表面分子标志CD73、CD90和CD105，不分泌M-CSF和EPO。UC-MSCs培养上清均能促进脐带血CD34~+细胞产生CFU-G、CFU-GM和CFU-M，但不产生CFU-E和CFU-GMEM。20~23岁供者UC-MSCs的增殖能力强于28~31岁供者。20~23岁生男孩的供者UC-MSCs细胞因子G-CSF、M-CSF、GM-CSF、IL-6、SOD2、TGF-β2、CDK6、JAG1的表达量较高，培养上清中促造血细胞因子GM-CSF、G-CSF和IL-6的含量较高，支持脐带血CD34+细胞形成较多集落。
[Abstract]:Umbilical cord mesenchymal stem cells (UC-MSCs) have the potential of self-renewal, proliferation and differentiation. They have important clinical application prospects in hematopoiesis recovery, repair of damaged or diseased tissues and organs, immunosuppressive therapy and so on. In order to obtain stable quality UC-MSCs, it is necessary to study the biological characteristics of UC-MSCs from different donors. Therefore, we studied the storage time of umbilical cord and the culture of UC-MSCs. We observed the cell morphology, proliferation, multidirectional differentiation, surface molecular markers and cytokine expression of UC-MSCs from different donor sources. The differences of their biological characteristics were compared. UC-MSCs culture supernatant was used to compare the ability of UC-MSCs from different donor sources to promote the secretion of hematopoietic factors. The storage time of fresh umbilical cord should not exceed 48 hours. When the umbilical cord was stored for 48 ~ 72 hours, the success rate of isolation of UC-MSCs decreased to 80%, the number of cells decreased to 3.83 脳 10 ~ 4 cm, the primary culture time extended to 10.8d.DMEM/F12 was the best medium for UC-MSCs, the primary culture time was 8.24 days, and the cell number was decreased to 3.83 脳 10 ~ 4 cm, the primary culture time was 8.24 days. UC-MSCs from different donors with cell count of 3.84 脳 10 脳 10, 6.20, and 31 years old were cultured in fusiform and adherent to each other. They were able to differentiate into adipocytes and osteoblasts in the induction medium, and expressed surface molecular markers CD73,CD90 and CD105,. Non-secreting M-CSF and EPO.UC-MSCs can promote the production of CFU-G,CFU-GM and CFU-M, in cord blood CD34~ cells. But non-CFU-E and CFU-GMEM.20~23-year-old donors UC-MSCs had higher proliferative capacity than 28-31-year-old donors. 20-23-year-old donors had UC-MSCs cytokines, including UC-MSCs cytokines, MMoCSF, GM CSF, IL-6, SOD2, TGF-尾 2, CDK6,. The expression of JAG1 was higher, and the contents of GM-CSF,G-CSF and IL-6 in the supernatant were higher, which supported the formation of more colonies of CD34 cells in cord blood.
【学位授予单位】：天津大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R329

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