论文部分内容阅读
采用0.25%胰蛋白酶与0.1%Ⅱ型胶原酶等体积混合灌流消化法培养人脐静脉内皮细胞(HUVEC),相差显微镜下,通过细胞形态学及Ⅷ因子免疫组织化学方法对培养的细胞进行鉴定,并测定不同配比的(5%、10%、15%、20%、30%)混合脐血浆(HUP)与20%胎牛血清(FBS)混搭对HUVEC增殖活性及细胞周期的影响。培养至7 d的HUVEC呈现出典型的铺路石状镶嵌生长,Ⅷ因子免疫组织化学染色阳性,20%HUP,15%HUP+20%FBS和20%HUP+20%FBS组较20%FBS对照组更有利于提高HUVEC的增殖活性。相反,30%HUP+20%FBS,则会引起细胞周期的阻滞,进而显著抑制HUVEC的增殖。结果证实,虽作用有限,尚不可完全替代血管内皮生长因子(VEGF),但作为培养液的添加成分之一,HUP不失为FBS的理想替代品。
Cultured human umbilical vein endothelial cells (HUVEC) were cultured in a volumetric mixed perfusion method with 0.25% trypsin and 0.1% collagenase type Ⅱ. The cultured cells were identified by cytomorphometry and Ⅷ factor immunohistochemical staining under a phase contrast microscope. The effects of different proportions (5%, 10%, 15%, 20%, 30%) of mixed Umbilical cord blood plasma (HUP) and 20% fetal bovine serum (FBS) on the proliferation and cell cycle of HUVEC were determined. HUVECs cultured for up to 7 days exhibited typical paving stone mosaic growth and were positive for factor Ⅷ immunohistochemical staining. Compared with 20% FBS control group, 20% HUP, 15% HUP + 20% FBS and 20% HUP + 20% More conducive to improve the proliferation of HUVEC activity. On the contrary, 30% HUP + 20% FBS, will lead to cell cycle arrest, thereby significantly inhibiting HUVEC proliferation. The results confirmed that, although limited, is not yet completely replace vascular endothelial growth factor (VEGF), but as an additional component of culture broth, HUP is an ideal substitute for FBS.