Expression of p75 neurotrophin receptor during the mineralization of ectomesenchymal stem cells in vitro
10.3760/cma.j.issn.1002-0098.2016.07.010
- VernacularTitle:p75神经营养受体在胎鼠颌突外胚间充质干细胞体外矿化过程中表达变化的研究
- Author:
Yingxin JU
1
;
Xiujie WEN
;
Kun YANG
;
Gang LI
;
Junyu LIU
;
Luchuan LIU
Author Information
1. 第三军医大学大坪医院野战外科研究所口腔科
- Keywords:
Mesenchymal stem cells;
Receptors,nerve growth factor;
Facial process;
Mineralization
- From:
Chinese Journal of Stomatology
2016;51(7):426-431
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the mineralized capacities of ectomesenchymal stem cells (EMSC) from facial process of Sprague Dawley(SD) rat embryo of different age in vitro and the expression of p75 neurotrophin receptor(p75NTR) in this process.Methods The stem cell surface antigens of EMSC from 12.5 d,15.5 d and 18.5 d SD rat embryonic facial process were tested by flow cytometry technology.E12.5 d EMSC,E15.5 d EMSC and E18.5 d EMSC were incubated under mineralization induction and analysed by alkaline phosphatase(ALP) staining on day 7(d7) and alizarin red staining on day 21(d21).Expression changes of Runt-related transcription factor-2(RUNX2),collagen Ⅰ (Col Ⅰ) and p75NTR in each group were measured using Western blotting and real time(RT)-PCR on day 0(d0),day 7(d7),day 14(d14) and day 21(d21).Results The expression of the special substances CD29,CD146 and p75NTR in E12.5 d EMSC,E15.5 d EMSC and E18.5 d EMSC were positive,and the expression of CD45 was negative.The expression level of p75NTR in E18.5 d EMSC(84.04％) was much higher than that of E12.5 d EMSC (22.53％) and E15.5 d EMSC(81.43％).The mineralized capacities of E18.5 d EMSC was stronger than that of E12.5 d EMSC and E15.5 d EMSC.The higher expression of RUNX2,Col Ⅰ in E18.5 d EMSC(RUNX2:1.92±0.20,Col Ⅰ:1.85±0.66) was found compared with E12.5 d EMSC(RUNX2:0.38±0.02,Col Ⅰ:0.33±0.94)and E15.5 d EMSC(RUNX2:0.72±0.22,Col Ⅰ:0.64±0.07)(P＜0.05),and p75NTR in the E18.5 d EMSC experimental group(E12.5 d:0.79±0.23,E15.5 d:0.84±0.29,E18.5 d:1.35±0.22) was significantly higher than the in control group(E12.5 d:0.42±0.12,E15.5 d:0.43±0.13,E18.5 d:0.48±0.15)(P＜0.05).RTPCR further proved the results of the Western blotting.Conclusions p75NTR participated in the mineralization differentiation of EMSC.E18.5 d EMSC had a higher expression of p75NTR and stronger mineralization capacity and was the ideal engineering seed cells.
