Induction differentiation of rabbit adipose‑derived stromal cells into insulin‑producing cells in vitro

Sun,Yu; Zhang,Mengchao; Ji,Shangwei; Liu,Lin

doi:10.3892/mmr.2015.4305

Induction differentiation of rabbit adipose‑derived stromal cells into insulin‑producing cells in vitro

Authors:
- Yu Sun
- Mengchao Zhang
- Shangwei Ji
- Lin Liu
View Affiliations

Affiliations: Department of Intervention, China‑Japan Union Hospital of Jilin University, Changchun, Jilin 130033, P.R. China, Department of Radiology, China‑Japan Union Hospital of Jilin University, Changchun, Jilin 130033, P.R. China, Department of Gastroenterology, China‑Japan Union Hospital of Jilin University, Changchun, Jilin 130033, P.R. China
Published online on: September 9, 2015 https://doi.org/10.3892/mmr.2015.4305
Pages: 6835-6840

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

Mesenchymal stem cells (MSCs) with the ability to differentiate into insulin‑producing cells (IPCs) have become the most promising means of therapy for diabetes mellitus. Adipose‑derived stromal cells (AdSCs), having similar characteristics to those of derived MSCs, are known to exhibit extensive proliferation potential and are able to undergo multi‑lineage differentiation. Whether AdSCs can differentiate into insulin‑producing cells (IPCs), however, has not been sufficiently elucidated. Therefore, the present study sought to investigate the in vitro differentiation of rabbit (r)AdSCs into IPCs, which may provide an abundant source of cells to treat diabetes. rADSCs were obtained from liposuction aspirates and then induced with glucagon‑like peptide‑1 and nicotinamide to differentiate into insulin‑secreting cells. Differentiation was evaluated by the analysis of morphology, dithizone (DTZ) staining, reverse transcription polymerase chain reaction (RT‑PCR), western blot analysis and a glucose challenge assay with detection of insulin secretion by ELISA. Morphological phase‑contrast microscopic observation revealed typical islet‑like cell clusters following 21 days of differentiation. DTZ staining also showed that differentiated cells were positive and undifferentiated cells were negative for insulin production. Furthermore, RT‑PCR analysis confirmed the mRNA expression of insulin, PDX1 and GLUT2 in differentiated cells. Western blot analysis showed that insulin was expressed by the differentiated cells. The glucose challenge assay showed that insulin secretion of the IPCs was in a glucose dependent manner. These findings implied that AdSCs are able to differentiate into IPC in vitro, and are therefore promising candidates for the treatment of diabetes.

View Figures

View References

1	Gao D, Xie J, Zhang J, Feng C, Yao B, Ma K, Li J, Wu X, Huang S and Fu X: MSC attenuate diabetes-induced functional impairment in adipocytes via secretion of insulin-like growth factor-1. Biochem Biophys Res Commun. 452:99–105. 2014. View Article : Google Scholar : PubMed/NCBI
2	American Diabetes A: Diagnosis and classification of diabetes mellitus. Diabetes Care. 36(Suppl 1): S67–S74. 2013. View Article : Google Scholar : PubMed/NCBI
3	Shaw JE, Sicree RA and Zimmet PZ: Global estimates of the prevalence of diabetes for 2010 and 2030. Diabetes Res Clin Pract. 87:4–14. 2010. View Article : Google Scholar
4	Chen LB, Jiang XB and Yang L: Differentiation of rat marrow mesenchymal stem cells into pancreatic islet beta-cells. World J Gastroenterol. 10:3016–3020. 2004. View Article : Google Scholar : PubMed/NCBI
5	Oh SH, Muzzonigro TM, Bae SH, LaPlante JM, Hatch HM and Petersen BE: Adult bone marrow-derived cells trans-differentiating into insulin-producing cells for the treatment of type I diabetes. Lab Invest. 84:607–617. 2004. View Article : Google Scholar : PubMed/NCBI
6	Vija L, Farge D, Gautier JF, Vexiau P, Dumitrache C, Bourgarit A, Verrecchia F and Larghero J: Mesenchymal stem cells: Stem cell therapy perspectives for type 1 diabetes. Diabetes Metab. 35:85–93. 2009. View Article : Google Scholar : PubMed/NCBI
7	Wu XH, Liu CP, Xu KF, Mao XD, Zhu J, Jiang JJ, Cui D, Zhang M, Xu Y and Liu C: Reversal of hyperglycemia in diabetic rats by portal vein transplantation of islet-like cells generated from bone marrow mesenchymal stem cells. World J Gastroenterol. 13:3342–3349. 2007. View Article : Google Scholar : PubMed/NCBI
8	Nejad-Dehbashi F, Hashemitabar M, Orazizadeh M, Bahramzadeh S, Shahhosseini Pourshoushtary E and Khorsandi L: The effects of exendine-4 on insulin producing cell differentiation from rat bone marrow-derived mesenchymal stem cells. Cell J. 16:187–194. 2014.PubMed/NCBI
9	Zhang YH, Wang HF, Liu W, Wei B, Bing LJ and Gao YM: Insulin-producing cells derived from rat bone marrow and their autologous transplantation in the duodenal wall for treating diabetes. Anat Rec (Hoboken). 292:728–735. 2009. View Article : Google Scholar
10	Gimble JM, Katz AJ and Bunnell BA: Adipose-derived stem cells for regenerative medicine. Circ Res. 100:1249–1260. 2007. View Article : Google Scholar : PubMed/NCBI
11	Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, Moorman MA, Simonetti DW, Craig S and Marshak DR: Multilineage potential of adult human mesenchymal stem cells. Science. 284:143–147. 1999. View Article : Google Scholar : PubMed/NCBI
12	Noël D, Caton D, Roche S, Bony C, Lehmann S, Casteilla L, Jorgensen C and Cousin B: Cell specific differences between human adipose-derived and mesenchymalstromal cells despite similar differentiation potentials. Exp Cell Res. 314:1575–1584. 2008. View Article : Google Scholar
13	Tyndall A, Walker UA, Cope A, Dazzi F, De Bari C, Fibbe W, Guiducci S, Jones S, Jorgensen C, Le Blanc K, et al: Immunomodulatory properties of mesenchymal stem cells: A review based on an interdisciplinary meeting held at the Kennedy Institute of Rheumatology Division, London, UK, 31 October 2005. Arthritis Res Ther. 9:3012007. View Article : Google Scholar : PubMed/NCBI
14	Krampera M, Pasini A, Pizzolo G, Cosmi L, Romagnani S and Annunziato F: Regenerative and immunomodulatory potential of mesenchymal stem cells. Curr Opin Pharmacol. 6:435–441. 2006. View Article : Google Scholar : PubMed/NCBI
15	Takahara K, Ii M, Inamoto T, Komura K, Ibuki N, Minami K, Uehara H, Hirano H, Nomi H, Kiyama S, et al: Adipose-derived stromal cells inhibit prostate cancer cell proliferation inducing apoptosis. Biochem Biophys Res Commun. 446:1102–1107. 2014. View Article : Google Scholar : PubMed/NCBI
16	Li L, Li F, Qi H, Feng G, Yuan K, Deng H and Zhou H: Coexpression of Pdx1 and betacellulin in mesenchymal stem cells could promote the differentiation of nestin-positive epithelium-like progenitors and pancreatic islet-like spheroids. Stem Cells and Dev. 17:815–823. 2008. View Article : Google Scholar
17	Bruin JE, Erener S, Vela J, Hu X, Johnson JD, Kurata HT, Lynn FC, Piret JM, Asadi A, Rezania A and Kieffer TJ: Characterization of polyhormonal insulin-producing cells derived in vitro from human embryonic stem cells. Stem Cell Res. 12:194–208. 2014. View Article : Google Scholar
18	Wei R, Yang J, Hou W, Liu G, Gao M, Zhang L, Wang H, Mao G, Gao H, Chen G and Hong T: Insulin-producing cells derived from human embryonic stem cells: Comparison of definitive endoderm- and nestin-positive progenitor-based differentiation strategies. PloS one. 8:e725132013. View Article : Google Scholar : PubMed/NCBI
19	Gao F, Wu DQ, Hu YH, Jin GX, Li GD, Sun TW and Li FJ: In vitro cultivation of islet-like cell clusters from human umbilical cord blood-derived mesenchymal stem cells. Transl Res. 151:293–302. 2008. View Article : Google Scholar : PubMed/NCBI
20	Chandra V, Swetha G, Muthyala S, Jaiswal AK, Bellare JR, Nair PD and Bhonde RR: Islet-like cell aggregates generated from human adipose tissue derived stem cells ameliorate experimental diabetes in mice. PloS one. 6:e206152011. View Article : Google Scholar : PubMed/NCBI
21	Zuk PA, Zhu M, Ashjian P, De Ugarte DA, Huang JI, Mizuno H, Alfonso ZC, Fraser JK, Benhaim P and Hedrick MH: Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell. 13:4279–4295. 2002. View Article : Google Scholar : PubMed/NCBI
22	Moshtagh PR, Emami SH and Sharifi AM: Differentiation of human adipose-derived mesenchymal stem cell into insulin-producing cells: An in vitro study. J Physiol Biochem. 69:451–458. 2013. View Article : Google Scholar
23	Bregenholt S, Møldrup A, Blume N, Karlsen AE, Nissen Friedrichsen B, Tornhave D, Knudsen LB and Petersen JS: The long-acting glucagon-like peptide-1 analogue, liraglutide, inhibits beta-cell apoptosis in vitro. Biochem Biophys Res Commun. 330:577–584. 2005. View Article : Google Scholar : PubMed/NCBI
24	Hui H, Nourparvar A, Zhao X and Perfetti R: Glucagon-like peptide-1 inhibits apoptosis of insulin-secreting cells via a cyclic 5′-adenosine monophosphate-dependent protein kinase A- and a phosphatidylinositol 3-kinase-dependent pathway. Endocrinology. 144:1444–1455. 2003. View Article : Google Scholar : PubMed/NCBI
25	Li LX, MacDonald PE, Ahn DS, Oudit GY, Backx PH and Brubaker PL: Role of phosphatidylinositol 3-kinasegamma in the beta-cell: Interactions with glucagon-like peptide-1. Endocrinology. 147:3318–3325. 2006. View Article : Google Scholar : PubMed/NCBI
26	Wang X, Zhou J, Doyle ME and Egan JM: Glucagon-like peptide-1 causes pancreatic duodenal homeobox-1 protein translocation from the cytoplasm to the nucleus of pancreatic beta-cells by a cyclic adenosine monophosphate/protein kinase A-dependent mechanism. Endocrinology. 142:1820–1827. 2001.PubMed/NCBI
27	Ji D, Li GY and Osborne NN: Nicotinamide attenuates retinal ischemia and light insults to neurones. Neurochem Int. 52:786–798. 2008. View Article : Google Scholar
28	Yang SJ, Choi JM, Kim L, Park SE, Rhee EJ, Lee WY, Oh KW, Park SW and Park CY: Nicotinamide improves glucose metabolism and affects the hepatic NAD-sirtuin pathway in a rodent model of obesity and type 2 diabetes. J Nutr Biochem. 25:66–72. 2014. View Article : Google Scholar