title: [Biological characteristics of mesenchymal stem cell and hematopoietic stem cell in the co-culture system].
abstract: The aim of the present study was to obtain the qualified hematopoietic stem/progenitor cells (HSC/HPC) and human umbilical cord-mesenchymal stem cells (MSC) in vitro in the co-culture system. Cord blood mononuclear cells were separated from umbilical cord blood by Ficoll lymphocyte separation medium, and then CD34+HSC was collected by MACS immunomagnetic beads. The selected CD34+HSC/HPC and MSC were transferred into culture flask. IMDM culture medium with 15% AB-type cord plasma supplemented with interleukin-3 (IL-3), IL-6, thrombopoietin (TPO), stem cell factor (SCF) and FMS-like tyrosine kinase 3 ligand (Flt-3L) factors were used as the co-culture system for the amplification of HSC/HPC and MSC. The cellular growth status and proliferation on day 6 and 10 after co-culture were observed by using inverted microscope. The percentage of positive expression of CD34 in HSC/HPC, as well as the percentages of positive expressions of CD105, CD90, CD73, CD45, CD34 and HLA-DR in the 4thgeneration MSC, was tested by flow cytometry. Semisolid colony culture was used to test the HSC/HPC colony forming ability. The osteogenic, chondrogenesis and adipogenic ability of the 4thgeneration MSC were assessed. The karyotype analysis of MSC was conducted by colchicines. The results demonstrated that the HSC/HPC of co-culture group showed higher ability of amplification, CFU-GM and higher CD34+percentage compared with the control group. The co-cultured MSC maintained the ability to differentiate into bone cells, fat cells and chondrocytes. And the karyotype stability of MSC remained normal. These results reveal that the appropriate co-culture system for MSC and HSC is developed, and via this co-culture system we could gain both two kinds of these cells. The MSCs under the co-culture system maintain the biological characteristics. The CFU-GM ability, cell counting and the flow cytometry results of HSC/HPC under the co-culture system are conform to the criterion, showing that the biological functions of HSC/HPC are maintained.
authors: Guangdong Cord Blood Bank, Guangzhou 510663, China. wwei@chinacord.org.
authors: Wei
authors: W
authors: Wei
authors: Guangdong Cord Blood Bank, Guangzhou 510663, China.
authors: Chao
authors: C
authors: Xu
authors: Guangdong Cord Blood Bank, Guangzhou 510663, China.
authors: Zhi-Yong
authors: ZY
authors: Ye
authors: Guangdong Cord Blood Bank, Guangzhou 510663, China.
authors: Xiao-Jun
authors: XJ
authors: Huang
authors: Guangdong Cord Blood Bank, Guangzhou 510663, China.
authors: Jia-En
authors: JE
authors: Yuan
authors: Guangdong Cord Blood Bank, Guangzhou 510663, China.
authors: Tian-Bao
authors: TB
authors: Ma
authors: Guangdong Cord Blood Bank, Guangzhou 510663, China.
authors: Han-Biao
authors: HB
authors: Lin
authors: Guangdong Cord Blood Bank, Guangzhou 510663, China.
authors: Xiu-Qiong
authors: XQ
authors: Chen
mesh_terms: D018952:Antigens, CD34; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D002478:Cells, Cultured; D018920:Coculture Techniques; D005312:Fetal Blood; D005434:Flow Cytometry; D006412:Hematopoietic Stem Cells; D006801:Humans; D007377:Interleukin-3; D015850:Interleukin-6; D059630:Mesenchymal Stem Cells; D019089:Stem Cell Factor; D014470:Umbilical Cord; D051941:fms-Like Tyrosine Kinase 3
chemical_list: D018952:Antigens, CD34; C508596:IL3 protein, human; C508600:IL6 protein, human; D007377:Interleukin-3; D015850:Interleukin-6; D019089:Stem Cell Factor; C497970:FLT3 protein, human; D051941:fms-Like Tyrosine Kinase 3