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. 2015 Mar 22:21:845-54.
doi: 10.12659/MSM.893144.

Proliferation and differentiation of rat osteoporosis mesenchymal stem cells (MSCs) after telomerase reverse transcriptase (TERT) transfection

Chao Li  1 , Guojun Wei  1 , Qun Gu  1 , Qiang Wang  2 , Shuqin Tao  1 , Liang Xu  1
Affiliations

Proliferation and differentiation of rat osteoporosis mesenchymal stem cells (MSCs) after telomerase reverse transcriptase (TERT) transfection

Chao Li et al. Med Sci Monit. .

Abstract

Background: The aim of this study was to determine whether MSC are excellent materials for MSCs transplantation in the treatment of osteoporosis.

Material and methods: We studied normal, osteoporosis, and TERT-transfected MSC from normal and osteoporosis rats to compare the proliferation and osteogenic differentiation using RT-PCR and Western blot by constructing an ovariectomized rat model of osteoporosis (OVX). The primary MSC from model rats were extracted and cultured to evaluate the proliferation and differentiation characteristics.

Results: MSCs of osteoporosis rats obviously decreased in proliferation ability and osteogenic differentiation compared to that of normal rats. In contrast, in TERT-transfected MSC, the proliferation and differentiation ability, and especially the ability of osteogenic differentiation, were significantly higher than in osteoporosis MSC.

Conclusions: TERT-transfected MSCs can help osteoporosis patients in whom MSC proliferation and osteogenic differentiation ability are weak, with an increase in both bone mass and bone density, becoming an effective material for autologous transplantation of MSCs in further treatment of osteoporosis. However, studies are still needed to prove the in vivo effect, biological safety, and molecular mechanism of TERT-osteoporosis treatment. Additionally, because the results are from an animal model, more research is needed in generalizing rat model findings to human osteoporosis patients.

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Figures

Figure 1
Figure 1
Bone histopathologic examination of the femurs of OVX and Sham rats. PS: (A) OVX group; (B) sham group.
Figure 2
Figure 2
The grouth curve of OVX MSCs and normal MSCs.
Figure 3
Figure 3
Differental gene expression between OVX and Sham MSCs.
Figure 4
Figure 4
(A, B) OVX and sham MSCs stained by Alizarin red after osteogenic differentiation.
Figure 5
Figure 5
Plasmid transfected 293T cells after 48 h. PS: (A) pLV.ExBi-CMV-TERT-IRES-eGFP; (B) pLV.ExSi. CMV-eGFP.
Figure 6
Figure 6
Lentiviral vector transfected osteoporosis MSCs. PS: (A) TERT-opMSCs; (B) GFP-opMSCs.
Figure 7
Figure 7
TERT expression differences in 3 groups examined by real time PCR.
Figure 8
Figure 8
TERT expression differences in 3 groups examined by Western blot.
Figure 9
Figure 9
Flow cytometry analysis of TERT-opMSCs.
Figure 10
Figure 10
The grouth curve of 3 groups.
Figure 11
Figure 11
TERT-opMSCs stained by Alizarin red after osteogenic differentiation culture.
See this image and copyright information in PMC

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