Kang Hongyan, Deng Xiaoyan
School of Biological Science & Medical Engineering, Beihang University, Beijing 100191, China.
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 2011 Feb;28(1):115-20.
This study was aimed to look for the potential application of human umbilical plasma (HUP) in the culture of human umbilical vein endothelial cells (HUVECs). The effect of HUP concentration in cell culture medium on cell proliferation activity and cell cycle was studied. HUVECs were obtained by digesting the umbilical cord with 0.25% trypsin mixed with the equal volume of 0.1% collagenase II, then were identified by morphology and factor VIII immunohistochemistry under phase contrast microscopy. The cells cultured after 7 days showed the typical cobblestone morphology with factor VIII immunohistochemical staining positive. The study showed that the groups of 20% (HUP), 15% HUP+20% Fetal Bovine Serum (FBS) and 20% HUP+20%FBS enhanced cell proliferation activity significantly when compared with the control group (20% FBS without HUP). On the contrary, 30% HUP+20% FBS caused cell cycle arrest, which significantly hindered the proliferation of HUVECs. The study proved that although HUP might not be able to completely replace the role of vascular endothelial growth factor in cell culture, as a supplement ingredient, it was an ideal candidate to replace FBS in culture medium.
本研究旨在探寻人脐血浆（HUP）在人脐静脉内皮细胞（HUVECs）培养中的潜在应用。研究了细胞培养基中HUP浓度对细胞增殖活性和细胞周期的影响。用0.25%胰蛋白酶与等体积的0.1%胶原酶II混合消化脐带获取HUVECs，然后在相差显微镜下通过形态学和因子VIII免疫组织化学进行鉴定。培养7天后的细胞呈现典型的鹅卵石样形态，因子VIII免疫组织化学染色呈阳性。研究表明，与对照组（不含HUP的20%胎牛血清（FBS））相比，20%（HUP）组、15% HUP + 20% FBS组和20% HUP + 20% FBS组显著增强了细胞增殖活性。相反，30% HUP + 20% FBS导致细胞周期停滞，显著阻碍了HUVECs的增殖。该研究证明，尽管HUP可能无法完全替代血管内皮生长因子在细胞培养中的作用，但作为一种补充成分，它是培养基中替代FBS的理想候选物。
