本文關(guān)鍵詞： 人臍血 間充質(zhì)干細(xì)胞 體外培養(yǎng) 分離 擴(kuò)增　出處：《廣州醫(yī)學(xué)院》2010年碩士論文　論文類型：學(xué)位論文

【摘要】：【目的】 1.探討人臍血來源的間充質(zhì)干細(xì)胞(Mesenchymal stem cells,MSCs)在體外分離培養(yǎng)與擴(kuò)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(xì)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(fù)合枸櫞酸鈉抗凝,用相對密度為1.077g/L的Ficoll淋巴細(xì)胞分離液分離臍血的單個(gè)核細(xì)胞(Mononuclear,MNC),分兩組,一組以20%胎牛血清濃度的培養(yǎng)基進(jìn)行培養(yǎng)和擴(kuò)增,一組以30%胎牛血清濃度的培養(yǎng)基進(jìn)行培養(yǎng)和擴(kuò)增,用流式細(xì)胞儀檢測人臍血MSCs的表面標(biāo)志。 【結(jié)果】 1.來源于人臍血的單個(gè)核細(xì)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(xì)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(xì)胞,主要表現(xiàn)為破骨樣細(xì)胞和間充質(zhì)樣細(xì)胞,經(jīng)傳3代后,可得純化擴(kuò)增的人臍血MSCs。流式細(xì)胞儀檢測結(jié)果顯示,人臍血MSCs不表達(dá)造血干細(xì)胞表面標(biāo)志的相關(guān)抗原CD14、淋巴細(xì)胞表面標(biāo)志的相關(guān)抗原CD19,強(qiáng)表達(dá)間充質(zhì)干細(xì)胞表面標(biāo)志的相關(guān)抗原CD105、CD44。 2.用高濃度胎牛血清培養(yǎng)基培養(yǎng)的單個(gè)核細(xì)胞形成的克隆數(shù)與低濃度胎牛血清培養(yǎng)的單個(gè)核細(xì)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(kuò)增,為人臍血MSCs的進(jìn)一步研究奠定基礎(chǔ)。 2. 30%胎牛血清濃度培養(yǎng)基的應(yīng)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(xué)位授予單位】：廣州醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：R329

【引證文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前1條

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

，

本文編號：1471740