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  本文關鍵詞： 人臍血 間充質(zhì)干細胞 體外培養(yǎng) 分離 擴增　出處：《廣州醫(yī)學院》2010年碩士論文　論文類型：學位論文

【摘要】：【目的】 1.探討人臍血來源的間充質(zhì)干細胞(Mesenchymal stem cells,MSCs)在體外分離培養(yǎng)與擴增的可行性。 2.探討不同培養(yǎng)基濃度下人臍血間充質(zhì)細胞(HMSCs)培養(yǎng)的差異。 【方法】 在無菌條件下收集正常足月胎兒的臍帶血,經(jīng)復合枸櫞酸鈉抗凝,用相對密度為1.077g/L的Ficoll淋巴細胞分離液分離臍血的單個核細胞(Mononuclear,MNC),分兩組,一組以20%胎牛血清濃度的培養(yǎng)基進行培養(yǎng)和擴增,一組以30%胎牛血清濃度的培養(yǎng)基進行培養(yǎng)和擴增,用流式細胞儀檢測人臍血MSCs的表面標志。 【結(jié)果】 1.來源于人臍血的單個核細胞接種于20%胎牛血清濃度的培養(yǎng)基中后,可產(chǎn)生貼壁細胞,但數(shù)量稀少不能傳代,接種于30%胎牛血清濃度的培養(yǎng)基中后,可產(chǎn)生較多的貼壁細胞,主要表現(xiàn)為破骨樣細胞和間充質(zhì)樣細胞,經(jīng)傳3代后,可得純化擴增的人臍血MSCs。流式細胞儀檢測結(jié)果顯示,人臍血MSCs不表達造血干細胞表面標志的相關抗原CD14、淋巴細胞表面標志的相關抗原CD19,強表達間充質(zhì)干細胞表面標志的相關抗原CD105、CD44。 2.用高濃度胎牛血清培養(yǎng)基培養(yǎng)的單個核細胞形成的克隆數(shù)與低濃度胎牛血清培養(yǎng)的單個核細胞形成的克隆數(shù)相比差異有顯著意義(P0.05)。 【結(jié)論】 1.來源于人臍血的MSCs在體外可以分離培養(yǎng)、擴增,為人臍血MSCs的進一步研究奠定基礎。 2. 30%胎牛血清濃度培養(yǎng)基的應用明顯提高人臍血MSCs培養(yǎng)的成功率。
[Abstract]:[purpose] 1. To investigate the feasibility of isolation, culture and expansion of mesenchymal stem cells derived from human umbilical cord blood in vitro. 2. To investigate the difference of human umbilical cord blood mesenchymal cells (HMSCs) culture in different culture medium. [methods] Umbilical cord blood of normal full-term fetus was collected under aseptic condition and anticoagulant with sodium citrate. Mononuclear mononuclear cells (MNCs) were isolated from umbilical cord blood with a relative density of 1.077g / L of Ficoll lymphocytes. The mononuclear cells were divided into two groups. One group was cultured and amplified on the medium of 20% fetal bovine serum concentration, another group was cultured and amplified by 30% fetal bovine serum concentration. The surface markers of human umbilical cord blood MSCs were detected by flow cytometry. [results] 1. Mononuclear cells derived from human umbilical cord blood could produce adherent cells after inoculating in the medium of 20% fetal bovine serum concentration, but the number of adherent cells could not be subcultured. After inoculating in the medium of 30% fetal bovine serum concentration, more adherent cells were produced, mainly osteoclast cells and mesenchymal cells. The purified human umbilical cord blood MSCs.FCM analysis showed that human umbilical cord blood MSCs did not express CD14 associated with hematopoietic stem cell surface markers. CD19, the antigen associated with lymphocyte surface markers, strongly expressed CD105 and CD44, which were associated with the surface markers of mesenchymal stem cells. 2. The clone number of mononuclear cells cultured in high concentration fetal bovine serum medium was significantly different from that in low concentration fetal bovine serum culture medium. [conclusion] 1. MSCs derived from human umbilical cord blood can be isolated, cultured and amplified in vitro, and the further study of MSCs in human umbilical cord blood lay a foundation. 2. The application of 30% fetal bovine serum concentration medium significantly improved the success rate of human umbilical cord blood MSCs culture.
【學位授予單位】：廣州醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2010
【分類號】：R329

【引證文獻】

相關碩士學位論文 前1條

1 魯俊山;補腎中藥聯(lián)合BMP-2對人臍血間充質(zhì)干細胞體外增殖及成骨活性影響的實驗研究[D];南京中醫(yī)藥大學;2012年

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本文編號：1471740