The Comparison Of Different Separation And Culture Ways Of Mononuclear Cells Derive From Human Umbilical Cord Blood

AIM: To explore a better isolation and culture method of mononuclear cells derived from human umbilical cord blood in vitro with different conditions. METHODS: The umbilical cord blood of healthy parturiens was taken from the department of obstetrics of First Affiliated Hospital of Xinjiang Medical University between May 2007 to March 2008.Several factors which influence the process of mononuclear cells in primary culture and passage stage were analyzed including different isolation methods, the time of the firstmedium changing, different culture mediums and different levels of fetal bovine serum. Mononuclear cells were identified using the surface marker by flow cytometry on the 30th of primary culture. Results: There were significant differences in the quantities of mononuclear cells derived from human umbilical cord blood among different isolation methods(P<0.01). The numbers of mononuclear cells from the hespan precipitation were more than that from density gradient centrifugation and immunomagnetic beads isolation which were respectively (15.23±4.30)×106/mL, (3.71±1.14)×106/mL, (0.066±0.027)×106/mL. But mononuclear cells growth condition was below the mark from the hespan precipitation. When the other conditions were the same, under those conditions including the first medium changing on the fifth or seventh day , low glucose DMEM or IMDE medium be helpfulto the growth of MNCs. The results of CD34 positive rate of density gradient centrifugation and immunomagnetic beads isolation by flow cytometry were respectively 10.1%, 0.5%. Conclusion: The hespan precipitation was an high- performance isolation method of MNCs derived from human umbilical cord blood .But MNCs growth condition was below the mark and need deep conform by experiment. The cells from CD133 immunomagnetic beads isolation were high purity quotient. You can choose the proper method according with different aims. The relative good methods of culture conditions of MNCs were the first medium changing on the fifth or seventh day , low glucose DMEM or IMDE medium . There were no different between 30% fetal bovine serum and 20% fetal bovine serum .