【摘要】：背景:研究表明,內(nèi)皮克隆形成細(xì)胞移植對缺血損傷組織的血管新生具有促進(jìn)作用。然而,這一作用是否與其旁分泌效應(yīng)有關(guān),仍有待進(jìn)一步研究證實(shí)。目的:檢測人臍血來源內(nèi)皮克隆形成細(xì)胞條件培養(yǎng)基中細(xì)胞因子分泌譜,并觀察內(nèi)皮克隆形成細(xì)胞條件培養(yǎng)基對人臍靜脈內(nèi)皮細(xì)胞增殖、遷移及成管能力的影響。方法:從人臍血中分離、培養(yǎng)內(nèi)皮克隆形成細(xì)胞,并進(jìn)行細(xì)胞鑒定。采用抗體芯片對無血清內(nèi)皮克隆形成細(xì)胞條件培養(yǎng)基中的細(xì)胞因子進(jìn)行檢測。采用無血清EBM-2培養(yǎng)基作為對照組,應(yīng)用CCK-8、細(xì)胞劃痕實(shí)驗(yàn)、Matrigel成管實(shí)驗(yàn)檢測內(nèi)皮克隆形成細(xì)胞條件培養(yǎng)基對人臍靜脈內(nèi)皮細(xì)胞增殖、遷移及成管能力的影響。結(jié)果與結(jié)論:(1)原代培養(yǎng)細(xì)胞形態(tài):培養(yǎng)的細(xì)胞呈"鋪路石"樣生長,表達(dá)CD34,KDR及CD144,不表達(dá)CD45及CD133,Dil-acLDL攝取及FITC-UEA-I結(jié)合雙陽性,在Matrigel基質(zhì)膠中可形成管腔樣結(jié)構(gòu),以上均符合內(nèi)皮克隆形成細(xì)胞的特征;(2)抗體芯片檢測:內(nèi)皮克隆形成細(xì)胞條件培養(yǎng)基高表達(dá)30種促血管新生因子;(3)CCK-8檢測:實(shí)驗(yàn)組人臍靜脈內(nèi)皮細(xì)胞在24,48,72 h增殖活性均高于對照組(P0.01);(4)細(xì)胞劃痕實(shí)驗(yàn)結(jié)果顯示,實(shí)驗(yàn)組人臍靜脈內(nèi)皮細(xì)胞遷移率在12,24 h均高于對照組(P0.01);(5)與對照組相比,實(shí)驗(yàn)組人臍靜脈內(nèi)皮細(xì)胞在Matrigel基質(zhì)膠上可形成更多的管狀結(jié)構(gòu)(P0.01)。(6)結(jié)果表明:內(nèi)皮克隆形成細(xì)胞能通過旁分泌效應(yīng)促進(jìn)成熟內(nèi)皮細(xì)胞的生物活性。
[Abstract]:Background: studies have shown that endothelial clone-forming cell transplantation can promote angiogenesis in ischemic tissue. However, whether this effect is related to paracrine effect needs further study. Aim: to detect the cytokine secretion profile in the conditioned medium of human umbilical cord blood derived endothelial clone forming cells, and to observe the effects of endothelial clone forming cell conditioned medium on the proliferation, migration and tubular ability of human umbilical vein endothelial cells (HUVECs). Methods: the cells were isolated from human umbilical cord blood, cultured and identified. The cytokines in serum-free endothelial clone forming cell conditioned medium were detected by antibody microarray. Serum-free EBM-2 medium was used as control group, CCK-8, cell scratch test and Matrigel tube forming test were used to detect the effect of endothelial clone forming cell conditioned medium on the proliferation, migration and tube-forming ability of human umbilical vein endothelial cells (HUVECs). Results & conclusion: (1) Primary culture cell morphology: the cultured cells grew like "paving stone", the expression of CD34,KDR and CD144, did not express CD45 and CD133,Dil-acLDL uptake, and FITC-UEA-I binding double positive. A tube-like structure could be formed in the Matrigel matrix gel, all of which were consistent with the characteristics of endothelial clone-forming cells. (2) Detection of antibody chip: endothelial clone formed cell culture medium with high expression of 30 proangiogenic factors; (3) CCK-8 assay: the proliferation activity of human umbilical vein endothelial cells in the experimental group was higher than that in the control group at 24,48,72 h (P0.01); (4) the results of cell scratch test showed that the migration rate of human umbilical vein endothelial cells in the experimental group was higher than that in the control group at 12 h and 24 h (P0.01). (5) compared with the control group, Experimental group of human umbilical vein endothelial cells could form more tubular structure on Matrigel matrix gel (P0.01). (6). The results showed that endothelial clone forming cells could promote the biological activity of mature endothelial cells through paracrine effect.
【作者單位】： 南方醫(yī)科大學(xué);解放軍廣州軍區(qū)廣州總醫(yī)院;廣東省中山市第二人民醫(yī)院;
【基金】：國家自然科學(xué)基金面上項(xiàng)目(81571910) 廣東省科技計(jì)劃項(xiàng)目(2014A020212256) 廣州市科技計(jì)劃項(xiàng)目(201607010394)~~
【分類號】：R329.2

【相似文獻(xiàn)】

相關(guān)期刊論文 前1條

1 于樹娜;王寶松;姜紅心;史才興;鄭潔;蔣吉英;;HepG2及其克隆形成細(xì)胞端粒酶活性的異質(zhì)性研究[J];中國組織化學(xué)與細(xì)胞化學(xué)雜志;2012年04期

，

本文編號：2455647