title: [Comparative study on combined culture of human placenta-derived mesenchymal stem cells and human umbilical vein endothelial cells from same and different individuals].
abstract: OBJECTIVE
  To investigate the protocols of combined culture of human placenta-derived mesenchymal stem cells (HPMSCs) and human umbilical vein endothelial cells (HUVECs) from the same and different individuals on collagen material,to provide the.
  
  
  METHODS
  Under voluntary contributions, HPMSCs were isolated and purified from human full-term placenta using collagenase IV digestion and lymphocyte separation medium, and confirmed by morphology methods and flow cytometry, and then passage 2 cells were cultured under condition of osteogenic induction. HUVECs were isolated from fresh human umbilical vein by collagenase I digestion and subcultured to purification, and cells were confirmed by immunocytochemical staining of von Willebrand factor (vWF). There were 2 groups for experiment. Passage 3 osteoblastic induced HPMSCs were co-cultured with HUVECs (1:1) from different individuals in group A and with HUVECs from the same individual in group B on collagen hydrogel. Confocal laser scanning microscope was used to observe the cellular behavior of the cell-collagen composites at 1, 3, 5, and 7 days after culturing.
  
  
  RESULTS
  Flow cytometry showed that HPMSCs were strongly positive for CD90 and CD29, but negative for CD31, CD45, and CD34. After induction, alizarin red, alkaline phosphatase, and collagenase I staining were positive. HUVECs displayed cobble-stone morphology and stained positively for endothelial cell marker vWF. The immunofluorescent staining of CD31 showed that HUVECs in the cell-collagen composite of group B had richer layers, adhered and extended faster and better in three-dimension space than that of group A. At 7 days, the class-like microvessel lengths and the network point numbers were (6.68 +/- 0.35) mm/mm2 and (17.10 +/- 1.10)/mm2 in group A, and were (8.11 +/- 0.62) mm/mm2 and (21.30 +/- 1.41)/mm2 in group B, showing significant differences between the 2 groups (t = 0.894, P = 0.000; t = 0.732, P = 0.000).
  
  
  CONCLUSIONS
  Composite implant HPMSCs and HUVECs from the same individual on collagen hydrogel is better than HPMSCs and HUVECs from different individuals in integrity and continuity of the network and angiogenesis.
authors.affiliation: Department of Orthopaedics, West China Hospital, Sichuan University, Chengdu Sichuan, 610041, P R China.
authors.forename: Jing
authors.initials: J
authors.lastname: Yang
authors.forename: Zhou
authors.initials: Z
authors.lastname: Xiang
authors.forename: Daiqing
authors.initials: D
authors.lastname: Wei
authors.forename: Xiao
authors.initials: X
authors.lastname: Huang
authors.forename: Yu
authors.initials: Y
authors.lastname: Gao
authors.forename: Bo
authors.initials: B
authors.lastname: Gao
authors.forename: Tao
authors.initials: T
authors.lastname: Shu
authors.forename: Xin
authors.initials: X
authors.lastname: Duan
authors.forename: Xingdong
authors.initials: X
authors.lastname: Zhang
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002478:Cells, Cultured; D018920:Coculture Techniques; D003094:Collagen / Q000378:metabolism; D005260:Female; D005434:Flow Cytometry; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology*; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D010012:Osteogenesis; D010920:Placenta / Q000166:cytology*; D011247:Pregnancy; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds
chemical_list: D003094:Collagen