Basic fibroblast growth factor lentiviral vector-transfected sheep bone marrow mesenchymal stem cells and non-specific osteogenic gene expression

Hu,Yang; Zhang,Qin; Zhang,Lei; Tang,Xiao‑Xue; He,Hui‑Yu

doi:10.3892/mmr.2015.3399

Basic fibroblast growth factor lentiviral vector-transfected sheep bone marrow mesenchymal stem cells and non-specific osteogenic gene expression

Authors:
- Yang Hu
- Qin Zhang
- Lei Zhang
- Xiao‑Xue Tang
- Hui‑Yu He
View Affiliations

Affiliations: Department of Prosthodontics, The First Affiliated Hospital of Xinjiang Medical University, Wulumuqi, Xinjiang 830054, P.R. China
Published online on: February 27, 2015 https://doi.org/10.3892/mmr.2015.3399
Pages: 267-272

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

In the present study, sheep bone marrow stem cells (BMSCs) were transfected with basic fibroblast growth factor (bFGF)‑lentivirus in order to investigate the influence of bFGF on osteogenic gene expression. bFGF‑transfected BMSCs (experimental group) and non‑transfected BMSCs (control group) were also transfected with a green fluorescent protein gene in order to measure the transfection efficiency of the (bFGF)‑lentivirus using flow cytometry. Using quantitative polymerase chain reaction assays, the changes in expression from three genes (osteopontin, OPN; osteocalcin, OC; and collagen‑I) in BMSCs from the experimental and control groups were measured. Transfection efficiency was 87.3% in the experimental group and 1.1% in the control group. OPN gene expression was high in BMSCs from the experimental group. However, there was no significant difference in OPN expression between BMSCs from the control and experimental group (P>0.05). Collagen‑Ⅰ expression was significantly lower in the experimental group compared with that in the control groups (P<0.05). By contrast, OC expression was significantly higher in BMSCs from the experimental group compared with that in the control group (P<0.05). Changes in osteogenic gene expression indicated that the BMSCs from the experimental group had better osteogenic ability, as compared with the control cells. Therefore, bFGF‑transfected cells may be useful seed cells for bone tissue engineering.

View Figures

View References

1	Bai Y, Li P, Yin G, et al: BMP-2, VEGF and bFGF synergistically promote the osteogenic differentiation of rat bone marrow-derived mesenchymal stem cells. Biotechnol Lett. 35:301–308. 2013. View Article : Google Scholar
2	Murakami S: Peridontal tissue regeneration by signaling molecule(s): what role does basic fibroblast growth factor (FGF-2) have in peridontal therapy? Peridontol 2000. 56:188–208. 2011. View Article : Google Scholar
3	Cockrell AS and Kafri T: Gene delivery by lentivirus vectors. Mol Biotechnol. 36:184–204. 2007. View Article : Google Scholar : PubMed/NCBI
4	Kotev-Emeth S, Pitary S, Pri-Chen S and Savion N: Establishment of a rat long-term culture expressing the osteogenic phenotype: dependence on dexamethasone and FGF-2. Connect Tissue Res. 43:606–612. 2002. View Article : Google Scholar
5	Yang Hu, Ying Ma and Huiyu He: The construction of eukaryotic expression lentiviral vector with basic fibroblast growth factor and transfection. Zhongguo zu zhi gong cheng yan jiu yu lin chuang kang fu za zhi. 15:1009–1014. 2011.In Chinese.
6	Farhadi J, Jaquiery C, Barbero A, et al: Differentiation-dependent up-regulation of BMP-2, TGF-beta1 and VEGF expression by FGF-2 in human bone marrow stromal cells. Plast Reconstr Surg. 116:1379–1386. 2005. View Article : Google Scholar : PubMed/NCBI
7	Wang L, Pang KF, Huang YL and Zhang XL: Experimental study of effects of different concentrations of bFGF proliferation and differentiation of canine bone marrow stromal cells. Kouqiang he mian wai ke za zhi. 18:6–10. 2008.In Chinese.
8	Locklin RM, Oreffo RO and Triffitt JJ: Effects of TGFbeta and bFGF on the differentiation of human bone marrow stromal fibroblasts. Cell Biol. 23:185–194. 1999.
9	Zhang JW, Feng JX, Xu RM, Zhao JZ and Zeng BF: Slow virus mediated alkaline fibroblast growth factor gene transfection meniscal fibrocartilage cells proliferation and matrix synthesis. Zhongguo zu zhi gong cheng yan jiu yu lin chuang kang fu za zhi. 11:8267–8270. 2007.In Chinese.
10	Indraccolo S, Habeler W, Tisato V, Stievano L, Piovan E, Tosello V, Esposito G, Wagner R, Uberla K, Chieco-Bianchi L and Amadori A: Gene transfer in ovarian cancer cells: a comparison between retroviral and lentivial vectors. Cancer Res. 62:6009–6107. 2002.
11	Wong LJ and Bai RK: Real-time quantitative polymerase chain reaction analysis of mitochondrial DNA point mutation. Methods In Mol Biol. 335:187–200. 2006.
12	Schmittgen TD: Real-time quantitative PCR. Methods. 25:383–385. 2001. View Article : Google Scholar
13	Liu XR, Zhang L and Wang YP: The theoretical research of real-time fluorescent quantitative PCR technology and medical applications. Zhongguo zu zhi gong cheng yan jiu yu lin chuang kang fu za zhi. 14:329–332. 2010.In Chinese.
14	Li YX, Li ZT and Wang YL: Real-time fluorescent quantitative PCR in the field of food testing applications. Liangshi yu you zhi za zhi. 2:38–40. 2010.In Chinese.
15	Chen X, Qi FK, Kang LG, et al: Real-time fluorescent quantitative research progress and application technology. Dongbei nong ye da xue xue bao za zhi. 41. pp. 148–155. 2010, In Chinese.
16	Tu LL, Zhang XL, Liu DQ, et al: In cotton fiber development and process of somatic embryogenesis in implementing control gene screening. Kexue tong bao za zhi. 52:2379–2385. 2007.In Chinese.
17	Yu SW, Liu HY and Luo LJ: Analysis of relative gene expression using different real-time quantitative PCR. Acta Agron Sin. 33:1214–1218. 2007.In Chinese.
18	Wei M, Xiong JH, Li YS and Fu BY: Study on glycogen synthase kinase gene expression variation under drought stress in rice by real-time PCR. Chinese Journal of Rice Science. 20:567–571. 2006.In Chinese.
19	Wang Q, Huang C, Zhang ZB, Hu JY and Zhang RZ: Real-time quantitative RTPCR method detection primitive culture medaka oestrogen induced fish liver cell gene expression. Huanjing ke xue xue bao za zhi. 28:2568–2572. 2008.In Chinese.
20	Zhang SB, Zhao YS, Liu X, Wang C and Liu GQ: Real-time quantitative PCR detection peas II etiolation seedling of actin isoforms expression. Beifang yuan yi za zhi. 110–113. 2010.In Chinese.
21	Wei XX, Li XZ, Tong GX and Wu XQ: Real-time fluorescent quantitative PCR technique and its application in aquatic animal viral disease quantitative detection. Shuichan ke xue za zhi. 29:681–686. 2010.In Chinese.
22	Kim HJ, Park HD, Kim JH, Cho JY, Choi JY, Kim JK, Kim HJ, Shin HI and Ryoo HM: Establishment and characterization of a stable cell line to evaluate cellular Runx2 activity. J Cel Biochem. 91:1239–1247. 2004. View Article : Google Scholar
23	Xiao G, Jiang D, Gopalakrishnan R and Franceschi RT: Fibroblastgrowth factor 2 induction of the osteocalcin gene requires MAPK activity and phosphorylation of the osteoblast transcription factor, Cbfa1/Runx2. J Biol Chem. 277:36181–36187. 2002. View Article : Google Scholar : PubMed/NCBI
24	Teplyuk NM, Haupt LM, Ling L, et al: The osteogenic transcription factor Runx2 regulates components of the fibroblast growth factor/proteoglycan signaling axis in osteoblasts. J Cell Biochem. 107:144–154. 2009. View Article : Google Scholar : PubMed/NCBI
25	Cotter EJ, Ip HS, Powderly WG and Doran PP: Mechanism of HIV protein induced modulation of mesenchymal stem cell osteogenic differentiation. BMC Musculoskelet Disord 9: 33. 2008. View Article : Google Scholar