人脐带间充质干细胞分泌肝细胞生长因子的初步研究  

A preliminary study of the level of hepatocyte growth factor secreted by human umbilical cord mesenchymal stem cell

作　　者：王荣[1,2] 邓运涛[2] 陈强[1,2] 

机构地区：[1]中国医学科学院北京协和医学院输血研究所,四川成都610081 [2]四川省脐带血造血干细胞库

出　　处：《中国输血杂志》2014年第8期825-829,共5页Chinese Journal of Blood Transfusion

摘　　要：目的研究体外传代培养的人脐带间充质干细胞（UC-MSCs）分泌肝细胞生长因子（HGF）的量及其变化。方法无菌条件下收集健康胎儿脐带15（人）份,并收集脐带供者资料,脐带采用组织块贴壁法分离培养UC-MSCs,并做传代培养;P3代UC-MSCs流式细胞仪检测免疫表型,成骨成脂诱导分化鉴定;采用ELISA法检测细胞培养上清中HGF的含量。结果 15份健康胎儿脐带均培养出UC-MSCs。P3代UC-MSCs的细胞表型均为CD90＋/CD105＋/CD166＋/CD14-/CD34-/CD45-/CD79a-/HLA-DR-。3份P3代细胞分别经过成骨和成脂诱导分化培养,诱导分化成骨细胞和脂肪细胞。体外传代培养的UC-MSCs均分泌细胞因子HGF;各传代细胞（n=15）P1、P3、P5和P7细胞上清中HGF含量（pg/mL）分别为32 754±15 147、26 029±11 680、11 660±5 137和3 105±2 197。15份脐带供者信息包括：孕产妇年龄（24~41岁）、孕周（37~40周）、胎儿体重（2.8~4.1kg）、胎儿性别（男性7例、女性8例）。上述供者因素与P3和P5代UC-MSCs分泌的HGF量无相关性（P〉0.05）。结论 UC-MSCs分泌HGF的量随着传代次数的增加而降低,且上述供者因素对UC-MSCs分泌HGF的量没有影响。Objective To study the levels and changes of the secretion of hepatocyte growth factor （HGF） from human umbilical cord mesenchymal stem cells （UC-MSCs） cultured in vitro. Methods The umbilical cords were collected from healthy fetal in sterile condition（ n = 15 ）. The donor information were collected. Mesenchymal stem cells were obtained by tis- sue adherence method and expanded. P3 cells were detected immunophenotype by flow cytometry, and induced osteogenic dif- ferentiation and adipogenic differentiation. We tested the level of HGF by ELISA method. Results UC-MSCs were all cul- tured from 15 healthly fetal. The P3 of UC-MSCs express CD90,CD105 and CD166,but no CD14,CD34,CD45 ,CD79a and HLA-DR. The P3 of UC-MSCs could be induced into adipocytes and osteoblasts（ n = 3 ）. HGF can be secreted by UC-MSCs cultured in vitro. The concentration of HGF in the supernatant of the P1 ,P3,P5 and P7 cells are 32 754 ± 15 147（pg/mL）, 26 029± 11 680（ pg/mL）, 11 660 ± 5 137 （pg/mL）, 3 105 ± 2 197 （pg/mL） （n = 15 ）. The levels of the secretion of HGF decreased gradually from P1 cells to P＇5 cells, and dropped sharply from P5 cells. The donor information included：maternal age （24 - 41 years old） ; gestational age （ 37- 40 weeks ） ; fetal weight （ 2. 8- 4. 1 kg） ; fetal gender （ 7 males, 8 females ）. In our studies, the HGF secretion of the P3 and P5 cells derived from different donors is not influenced by donors ages, gestation- al age,delivery modes,fetal weight and fetal gender. Conclusion The level of the secretion of HGF decreased during pas- sage, and the donor factors had no effect on the concentration of HGF secreted by UC-MSCs.

关 键 词：人脐带间充质干细胞 肝细胞生长因子 组织块贴壁法 细胞培养 细胞因子 产妇年龄 孕周 胎儿体重 胎儿性别 

分 类 号：R457.7[医药卫生—治疗学] R329.26[医药卫生—临床医学]