Different culture methods for human umbilical cord mesenchymal stem cells / 中国组织工程研究

ABSTRACT

BACKGROUND:Umbilical cord stem cel s mainly derive from ful-term infants, and common culture methods include tissue-attached method and trypsin-digestion mehod. However, effects of different culture methods on the separation of umbilical cord mesenchymal stem cel s remain many disputes. OBJECTIVE:To observe the effects of different culture methods on umbilical cord mesenchymal stem cel s. METHODS:Umbilical cords of 30 healthy ful-term and caesarean delivery infants were selected, and cultured using tissue-attached method or trypsin-digestion method to isloate and culture human umbilical cord mesenchymal stem cel s. Meanwhile, cel growth was measured by flow cytometry. RESULTS AND CONCLUSION:The fusiform-shaped cel s began to separate from the umbilical cord tissue that was primary cultured using tissue-attached method, and 10 days later, the cel fusion reached 80%;after the umbilical cord was cultured using col agenase-trypsin digestion for 5 days, a smal amount of adherent cel s with different shapes appeared, and the fiber-like cel s reached 80%of confluence until 2-week culture. There was no significant difference in the growth of umbilical cord mesenchymal stem cel s cultured by different culture methods (P>0.05). Moreover, cel s cultured by two methods were al positive for CD13, CD29, CD44 and CD105. These results demonstrate that human umbilical cord mesenchymal stem cel s exhibit a high success rate in primary culture using tissue-attached method, which is superior to the trypsin-digestion method.