The Modified Culture Method Of Mesenchymal Stem Cell

PartⅠThe modified culture method of human umbilical cord mesenchymalstem cellObjective, To establish the method to isolate and culture human umbilical cordmesenchymal stem cells (hUCMSCs) in vitro.Methods, Mesenchymal stem cells were isolated from human umbilical cord byenzyme digestion, traditionary tissue adherence and modified tissue adherence, andcompared the growth status and quantity of their primary cells. The immunophenotypeof the cells cultivated by modified method were detected by flow cytometer. The cellswere induced to osteoblasts and lipoblasts by different culture systems. After induction,the cells were identified by specific methods.Results, The modified culture method of hUCMSCs was established. The primary cellnumber is nearly2times than got through enzyme digestion method and is about20times than traditionary method, with higher purity. The adherent cells cultivated bymodified method showed spindle shape under an inverted microscope and werepositive for CD90、CD44, but negative for CD45、CD34. After induction, alizarin redstaining and oil red O staining were positive.Conclusion, An effectively new method for hUCMSCs isolation and culture wasestablished, the potent expansion capacity and multiple differentiation potential can beconfirmed, which can be used to guide cell based therapies.PartⅡThe modified culture method of BALB/C mice bone marrow mesenchymalstem cellsObjective, To establish the method to isolate and culture BALB/C mice bone marrowmesenchymal stem cells (BMSCs) in vitro. Methods, Mesenchymal stem cells were isolated from mice bone marrow by thetraditionary whole bone marrow adherent culture, and the modified whole bone marrowadherent culture, and compared the growth status and quantity of their primary cells; Theimmunophenotype of the cells cultivated by modified method were detected by flowcytometer and the cells were induced to osteoblasts by osteoblast induced liquid. Afterinduction, the cells were identified by specific methods.Results, The primary cell number cultured by the modified whole bone marrow culturemethod increased50%than the traditional method, and the cells grow faster; After Fluidcytology testing, the adherent cells were positive for CD90、CD44, but negative for CD45;After induction, alizarin red staining were positive.Conclusion, An effectively new method for BALB/C mice bone marrow mesenchymalstem cell isolation and culture was established, which can be used to guide cell basedtherapies.