【摘要】：臍帶間充質(zhì)干細胞(UC-MSCs)具有自我更新、增殖和分化的潛能，在造血恢復、修復損傷或病變的組織器官、免疫抑制治療等方面具有重要臨床應用前景。為了獲得穩(wěn)定質(zhì)量的UC-MSCs，研究不同供者UC-MSCs生物學特性的差異是十分必要的。為此，我們對臍帶的存放時間和UC-MSCs的培養(yǎng)進行研究，通過觀察不同供者來源的UC-MSCs的細胞形態(tài)、增殖能力、多向分化能力、表面分子標志物和細胞因子的表達量，比較其生物學特性的差異。用UC-MSCs培養(yǎng)上清，比較不同供者來源UC-MSCs促造血因子的分泌能力。 采集后的新鮮臍帶存放時間不宜超過48h。臍帶存放時間48~72h時，分離UC-MSCs的成功率下降為80%，細胞數(shù)減少為3.83×10~4/cm、原代培養(yǎng)時間延長為10.8d。DMEM/F12是UC-MSCs的最適培養(yǎng)基，原代培養(yǎng)時間為8.24d，獲得的細胞數(shù)達3.84×10~6。20~31歲的不同供者的UC-MSCs體外培養(yǎng)均呈紡錘形、貼壁生長，在誘導培養(yǎng)基中均能分化為脂肪細胞和成骨細胞，表達表面分子標志CD73、CD90和CD105，不分泌M-CSF和EPO。UC-MSCs培養(yǎng)上清均能促進臍帶血CD34~+細胞產(chǎn)生CFU-G、CFU-GM和CFU-M，但不產(chǎn)生CFU-E和CFU-GMEM。20~23歲供者UC-MSCs的增殖能力強于28~31歲供者。20~23歲生男孩的供者UC-MSCs細胞因子G-CSF、M-CSF、GM-CSF、IL-6、SOD2、TGF-β2、CDK6、JAG1的表達量較高，培養(yǎng)上清中促造血細胞因子GM-CSF、G-CSF和IL-6的含量較高，支持臍帶血CD34+細胞形成較多集落。
[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.
【學位授予單位】：天津大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R329

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