Growth and expression of rat bone marrow mesenchymal stem cells modified by nerve growth factor in diabetic rat bladders

Zhu,Shao‑Xing; Huang,Shi‑Yong; Su,Yi‑Ming; Cai,Peng

doi:10.3892/mmr.2013.1425

Growth and expression of rat bone marrow mesenchymal stem cells modified by nerve growth factor in diabetic rat bladders

Authors:
- Shao‑Xing Zhu
- Shi‑Yong Huang
- Yi‑Ming Su
- Peng Cai
View Affiliations

Affiliations: Department of Urology, Zhejiang Cancer Hospital, Hangzhou 310022, P.R. China, Department of Urology, Xiamen Hospital of Traditional Chinese Medicine, Xiamen 361000, P.R. China, Department of Urology, Union Hospital, Fujian Medical University, Fuzhou 350001, P.R. China, Department of Urology, Xiangcheng Hospital, Zhangzhou 363000, P.R. China
Published online on: April 10, 2013 https://doi.org/10.3892/mmr.2013.1425
Pages: 1791-1799

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

The aim of the present study was to determine whether rat bone marrow mesenchymal stem cells (MSCs) transfected with the nerve growth factor (NGF) gene and then transplanted into diabetic rat bladder tissues survive and continue to express NGF. A recombinant lentiviral vector carrying the NGF gene was constructed and transfected into rat bone marrow MSCs. BrdU‑labeled immunohistochemistry was used to observe NGF expression in the transfected MSCs. BrdU‑labeled and NGF‑transfected MSCs were transplanted into diabetic rat bladder tissues. BrdU‑labeled immunohistochemistry was used to observe the growth of NGF‑transfected MSCs in the tissue samples. NGF mRNA and protein expression levels in MSCs were analyzed using reverse transcription polymerase chain reaction (RT-PCR) and ELISA, respectively. The recombinant NGF gene lentiviral vector and NGF gene-modified rat bone marrow MSCs were successfully constructed. NGF gene-modified rat MSCs survived in the diabetic rat bladders 4 weeks following injection and NGF gene expression was increased. In the present study, NGF gene-modified MSCs were shown to be capable of survival in diabetic rat bladder tissues and stably expressed NGF.

View Figures

View References

1	Ueda T, Yoshimura N and Yoshida O: Diabetic cystopathy: relationship to autonomic neuropathy detected by sympathetic skin response. J Urol. 157:580–584. 1997. View Article : Google Scholar : PubMed/NCBI
2	Paro M, Italiano G, Travagli RA, et al: Cystometric changes in alloxan diabetic rats: evidence for functional and structural correlates of diabetic autonomic neuropathy. J Auton Nerv Syst. 30:1–11. 1990. View Article : Google Scholar : PubMed/NCBI
3	Hellweg R, Raivich G, Hartung HD, Hock C and Kreutzberg GW: Axonal transport of endogenous nerve growth factor (NGF) and NGF receptor in experimental diabetic neuropathy. Exp Neurol. 130:24–30. 1994. View Article : Google Scholar : PubMed/NCBI
4	Sasaki K, Chancellor MB, Phelan MW, et al: Diabetic cystopathy correlates with a long-term decrease in nerve growth factor levels in the bladder and lumbosacral dorsal root ganglia. J Urol. 168:1259–1264. 2002. View Article : Google Scholar : PubMed/NCBI
5	Goins WF, Yoshimura N, Phelan MW, et al: Herpes simplex virus mediated nerve growth factor expression in bladder and afferent neurons: potential treatment for diabetic bladder dysfunction. J Urol. 165:1748–1754. 2001. View Article : Google Scholar
6	Sasaki K, Chancellor MB, Goins WF, et al: Gene therapy using replication-defective herpes simplex virus vectors expressing nerve growth factor in a rat model of diabetic cystopathy. Diabetes. 53:2723–2730. 2004. View Article : Google Scholar
7	Yokoyama T, Yoshimura N, Dhir R, et al: Persistence and survival of autologous muscle derived cells versus bovine collagen as potential treatment of stress urinary incontinence. J Urol. 165:271–276. 2001. View Article : Google Scholar
8	Strasser H, Marksteiner R, Margreiter E, et al: Transurethral ultrasonography-guided injection of adult autologous stem cells versus transurethral endoscopic injection of collagen in treatment of urinary incontinence. World J Urol. 25:385–392. 2007. View Article : Google Scholar
9	Huard J, Yokoyama T, Pruchnic R, et al: Muscle-derived cell-mediated ex vivo gene therapy for urological dysfunction. Gene Ther. 9:1617–1626. 2002. View Article : Google Scholar : PubMed/NCBI
10	Pedersini R, Vattemi E and Claudio PP: Adenoviral gene therapy in high-grade malignant glioma. Drug News Perspect. 23:368–379. 2010.PubMed/NCBI
11	Dreyer JL: Lentiviral vector-mediated gene transfer and rna silencing technology in neuronal dysfunctions. Mol Biotechnol. 47:169–187. 2011. View Article : Google Scholar : PubMed/NCBI
12	Benoit RM, Wilhelm RN, Scherer-Becker D and Ostermeier C: An improved method for fast, robust, and seamless integration of DNA fragments into multiple plasmids. Protein Expr Purif. 45:66–71. 2006. View Article : Google Scholar : PubMed/NCBI
13	Berrow NS, Alderton D, Sainsbury S, et al: A versatile ligation-independent cloning method suitable for high-throughput expression screening applications. Nucleic Acids Res. 35:e452007. View Article : Google Scholar
14	Klock HE, White A, Koesema E and Lesley SA: Methods and results for semi-automated cloning using integrated robotics. J Struct Funct Genomics. 6:89–94. 2005. View Article : Google Scholar : PubMed/NCBI
15	Friedenstein AJ, Petrakova KV, Kurolesova AI and Frolova GP: Heterotopic of bone marrow. Analysis of precursor cells for osteogenic and hematopoietic tissues. Transplantation. 6:230–247. 1968.PubMed/NCBI
16	Porada CD, Zanjani ED and Almeida-Porad G: Adult mesenchymal stem cells: a pluripotent population with multiple applications. Curr Stem Cell Res Ther. 1:365–369. 2006. View Article : Google Scholar : PubMed/NCBI
17	Hristov M, Heussen N, Schober A and Weber C: Intracoronary infusion of autologous bone marrow cells and left ventricular function after acute myocardial infarction: a meta-analysis. J Cell Mol Med. 10:727–733. 2006. View Article : Google Scholar : PubMed/NCBI
18	Kanazawa H, Fujimoto Y, Teratani T, et al: Bone marrow-derived mesenchymal stem cells ameliorate hepatic ischemia reperfusion injury in a rat model. PLoS One. 6:e191952011. View Article : Google Scholar : PubMed/NCBI
19	Choi H, Lee RH, Bazhanov N, Oh JY and Prockop DJ: Anti-inflammatory protein TSG-6 secreted by activated MSCs attenuates zymosan-induced mouse peritonitis by decreasing TLR2/NF-κB signaling in resident macrophages. Blood. 118:330–338. 2011.PubMed/NCBI
20	Xu YL, Liu YL, Wang Q, Li G, Lü XD and Kong B: Intravenous transplantation of mesenchymal stem cells attenuates oleic acid induced acute lung injury in rats. Chin Med J (Engl). 125:2012–2018. 2012.PubMed/NCBI