[Related factors affecting the isolation of multipotent non-hematopoietic adult stem cells from umbilical cord blood].
To investigate the related factors affecting the isolation of multipotent non-hematopoietic adult stem cells (MNASCs) from human umbilical cord blood in low serum (2%) condition, the isolation conditions were optimized and the yield of MNASCs was improved. MNASCs from human umbilical cord blood samples were isolated, and the effects of medium component, medium exchange time and initial plating density for isolation of MNASCs were studied. Then, the MNASCs were isolated and cultured in optimal condition, the surface antigen expression and differentiation potential of MNASCs were detected. The result showed that the medium of DMEM/F12 was better than IMDM and DMEM-LG for MNASCs culture in low serum condition. The optimal yield of MNASCs was obtained when mononuclear cells were cultured at a initial plating density of 1 x 10(6) cells/cm2 and the medium was exchanged to remove the nonadherent cells after 72 hours of inoculation. MNASCs isolated and cultured under the above-mentioned conditions maintained a homogenous morphology, high potential ability of expansion and differentiation. It is concluded that culture conditions with low serum defined in this study is optimal for the successful isolation and expansion of umbilical cord blood MNASCs with high numbers for subsequent cellular therapeutic approaches.
Li
Yun-Tao
YT
State Key Laboratory of Experimental Hematology, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences and Peking Union Medical University, Tianjin 300020, China.
Xu
Yan
Y
Meng
Hen-Xing
HX
Ge
Wei
W
Li
Qiao-Chuan
QC
Wan
Chang-Chun
CC
Yu
Zhen
Z
Li
Chang-Hong
CH
Qiu
Lu-Gui
LG
D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000502:physiology; D002469:Cell Separation / Q000379:methods*; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000502:physiology; D005312:Fetal Blood / Q000166:cytology*; D006801:Humans; D039902:Multipotent Stem Cells / Q000166:cytology* / Q000502:physiology