Korean J Physiol Pharmacol.  2016 Jul;20(4):341-346. 10.4196/kjpp.2016.20.4.341.

The effect of progesterone and 17-β estradiol on membrane-bound HLA-G in adipose derived stem cells

• Moslehi A
• Hashemi-beni B
• Moslehi A
• Akbari MA
• Adib M

• KMID: 2371051
• DOI: http://doi.org/10.4196/kjpp.2016.20.4.341

Abstract

Membrane-bound HLA-G (mHLA-G) discovery on adipose derived stem cells (ADSCs) as a tolerogenic and immunosuppressive molecule was very important. Many documents have shown that HLA-G expression can be controlled via some hormones such as progesterone (P4) and estradiol (E2). Therefore, this study was designed to evaluate progesterone and estradiol effects on mHLA-G in ADSCs at restricted and combination concentrations. Three independent cell lines were cultured in complete free phenol red DMEM and subcultured to achieve suffi cient cells. These cells were treated with P4, E2 and P4 plus E2 at physiologic and pregnancy concentrations for 3 days in cell culture conditions. The HLA-G positive ADSCs was measured via monoclonal anti HLA-G-FITC/MEMG-09 by means of flow cytometry in nine groups. Data were analyzed by one way ANOVA and Tukey's post hoc tests. There were no signifi cant values of the mean percentage of HLA-G positive cells in E2-treated and the combination of P4 plus E2-treated ADSCs compared to control cells (p value>0.05) but P4 had a signifi cant increase on mHLA-G in ADSCs (p value<0.05). High P4 concentration increased mHLA-G but E2 and the combination of P4 plus E2 could not change mHLA-G on ADSCs.

Keyword

Adipose derived stem cell; Estradiol; Membrane-bound HLA-G; Progesterone

MeSH Terms

Cell Culture Techniques
Cell Line
Estradiol*
Flow Cytometry
HLA-G Antigens*
Phenolsulfonphthalein
Pregnancy
Progesterone*
Stem Cells*
Estradiol
HLA-G Antigens
Phenolsulfonphthalein
Progesterone

Figure

• 

  Fig. 1 Flow cytometry dot plot of HLA-G positive ADSCs in the absence of progesterone and estradiol.(A) Isotype control. (B) Control group. Shown percentages are the mean values of three independent experiments.

• 

  Fig. 2 Flow cytometry dot plot of HLA-G positive ADSCs in the presence of progesterone.(A) 1×10–5 M P4, (B) 3×10–5 M P4. (C) Statistical analysis of the percentage of HLA-G positive P4-treated cells compared to control cell. Shown percentages are the mean values of three independent experiments. ***<0.001; ns, not significant; c, control cell.

• 

  Fig. 3 Flow cytometry dot plot of HLA-G positive ADSCs in the presence of estradiol.(A) 1×10–9 M E2, (B) 1×10–7 M E2. (C) Statistical analysis of the percentage of HLA-G positive E2-treated cells compared to control cell. Shown percentages are the mean values of three independent experiments (p>0.05). ns, not significant; c, control cell.

• 

  Fig. 4 Flow cytometry dot plot of HLA-G positive ADSCs in the presence of P4 plus E2.Panels (A) E2: 10–7 M+P4:1×10–5 M, (B) E2: 10–7 M+P4:3×10–5 M, (C) E2:10–9 M+P4:1×10–5 M, (D) E2:10–9 M+ P4:3×10–5 M. (E) Statistical analysis of the percentage of HLA-G positive P4 plus E2-treated cells compared to P4-treated cells as control group. Shown percentages are the mean values of three independent experiments (p>0.05). ns, not significant. C1: 1×10–5 M P4, C2: 3×10–5 M P4.

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