Influence of proliferation and ostogenic differentiation of 1,25 (OH)2VD3 on hUCMSCs in vitro

Abstract

Objective To establish the separation and expansion method of human umbilical cord derived mesenchymal stem cells (hUCMSCs) and explore the influence of 1,25 (OH)2VD3 on hUCMSCs in vitro.Methods UCMSCs were cultured from adherent tissue pieces and detected by immunohistochemistry.Taking the third generation with good growth state UCMSCs make intervene experiment.Set 10-7 mol/L,10-9mol/L and 10-11 mol/L 3 kind of concentration of 1,25(OH)2VD3 as the experimental group,and a control group (DMEM medium).The morphologic change of UCMSCs was observed by inverted microscope.Observe the cell proliferation by the method of MTT.Detect alkaline phosphatase assay and type Ⅰ collagen assay from immunohistochemistry.And the calcium tubercle formation would be examined after 21 days.Results UCMSCs were cultured from adherent tissue pieces and strongly positive for CD44,CD105 and negative for CD34,CD45.consistent with the hUCMSCs characteristics.Under different concentration of 1,25 (OH)2VD3hUCMSCs proliferation were promoted within 7 days but were suppressed beyond 7 days.The high doses group ( 10-7 mol/L group ) obvious inhibitted the stem cell proliferation.Different concentrations of 1,25 (OH)2VD3 and days have interactive effect (P ＜ 0.05).The differences between different groups with ALP and type Ⅰ collagen assay has statistical difference (P ＜ 0.05).The secretion of ALP and type Ⅰ collagen assay of 10-7 mol/Lconcentration group was higher than others.Typical mineralization nedus was found in intervene groups.Conclusion HUCMSCs can be successfully cultured from the adherent tissue pieces.1,25(OH)2VD3 can effectively induce hUCMSCs to differentiate into osteoblasts in vitro. Key words: Umbilical cord derived mesenchymal stem cells (UCMSCs); 1,25(OH)2VD3; Differentiation; Osteoblasts