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Generation of high-yield insulin producing cells from human bone marrow mesenchymal stem cells


Allogenic islet transplantation is a most efficient approach for treatment of diabetes mellitus. However, the scarcity of islets and long term need for an immunosuppressant limits its application. Recently, cell replacement therapies that generate of unlimited sources of β cells have been developed to overcome these limitations. In this study we have described a stage specific differentiation protocol for the generation of insulin producing islet-like clusters from human bone marrow mesenchymal stem cells (hBM-MSCs). This specific stepwise protocol induced differentiation of hMSCs into definitive endoderm, pancreatic endoderm and pancreatic endocrine cells that expressed of sox17, foxa2, pdx1, ngn3, nkx2.2, insulin, glucagon, somatostatin, pancreatic polypeptide, and glut2 transcripts respectively. In addition, immunocytochemical analysis confirmed protein expression of the above mentioned genes. Western blot analysis discriminated insulin from proinsulin in the final differentiated cells. In derived insulin producing cells (IPCs), secreted insulin and C-peptide was in a glucose dependent manner. We have developed a protocol that generates effective high-yield human IPCs from hBM-MSCs in vitro. These finding suggest that functional IPCs generated by this procedure can be used as a cell-based approach for insulin dependent diabetes mellitus.

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  1. 1.

    Chen LB, Jiang XB, Yang L (2004) Differentiation of rat marrow mesenchymal stem cells into pancreatic islet beta-cells. World J Gastroenterol 10(20):3016–3020

    CAS  PubMed  Google Scholar 

  2. 2.

    Shim J, Kim S, Woo D, Kim S, Oh C, McKay R, Kim J (2007) Directed differentiation of human embryonic stem cells towards a pancreatic cell fate. Diabetologia 50(6):1228–1238

    CAS  Article  PubMed  Google Scholar 

  3. 3.

    Zhang D, Jiang W, Liu M, Sui X, Yin X, Chen S, Shi Y, Deng H (2009) Highly efficient differentiation of human ES cells and iPS cells into mature pancreatic insulin-producing cells. Cell Res 19(4):429–438

    CAS  Article  PubMed  Google Scholar 

  4. 4.

    Gao F, Wu D-Q, Hu Y-H, Jin G-X, Li G-D, Sun T-W, Li F-J (2008) In vitro cultivation of islet-like cell clusters from human umbilical cord blood-derived mesenchymal stem cells. Trans Res 151(6):293–302

    CAS  Article  Google Scholar 

  5. 5.

    Chao KC, Chao KF, Fu YS, Liu SH (2008) Islet-like clusters derived from mesenchymal stem cells in Wharton’s Jelly of the human umbilical cord for transplantation to control type 1 diabetes. PLoS ONE 3(1):e1451

    PubMed Central  Article  PubMed  Google Scholar 

  6. 6.

    Chang C-M, Kao C-L, Chang Y-L, Yang M-J, Chen Y-C, Sung B-L, Tsai T-H, Chao K-C, Chiou S-H, Ku H-H (2007) Placenta-derived multipotent stem cells induced to differentiate into insulin-positive cells. Biochem Biophys Res Commun 357(2):414–420

    CAS  Article  PubMed  Google Scholar 

  7. 7.

    Okura H, Komoda H, Fumimoto Y, Lee C-M, Nishida T, Sawa Y, Matsuyama A (2009) Transdifferentiation of human adipose tissue-derived stromal cells into insulin-producing clusters. J Artif Organs 12(2):123–130

    CAS  Article  PubMed  Google Scholar 

  8. 8.

    Colter DC, Class R, DiGirolamo CM, Prockop DJ (2000) Rapid expansion of recycling stem cells in cultures of plastic-adherent cells from human bone marrow. Proc Natl Acad Sci 97(7):3213–3218

    PubMed Central  CAS  Article  PubMed  Google Scholar 

  9. 9.

    Shiroi A, Yoshikawa M, Yokota H, Fukui H, Ishizaka S, Tatsumi K, Takahashi Y (2002) Identification of insulin-producing cells derived from embryonic stem cells by zinc-chelating dithizone. Stem Cells 20(4):284–292

    CAS  Article  PubMed  Google Scholar 

  10. 10.

    Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65(1):55–63

    CAS  Article  PubMed  Google Scholar 

  11. 11.

    Li Y, Zhang R, Qiao H, Zhang H, Wang Y, Yuan H, Liu Q, Liu D, Chen L, Pei X (2007) Generation of insulin-producing cells from PDX-1 gene-modified human mesenchymal stem cells. J Cell Physiol 211(1):36–44

    CAS  Article  PubMed  Google Scholar 

  12. 12.

    Chandra V, Swetha G, Muthyala S, Jaiswal AK, Bellare JR, Nair PD, Bhonde RR (2011) Islet-like cell aggregates generated from human adipose tissue derived stem cells ameliorate experimental diabetes in mice. PLoS ONE 6(6):e20615

    PubMed Central  CAS  Article  PubMed  Google Scholar 

  13. 13.

    Zhang N, Li J, Luo R, Jiang J, Wang J-A (2008) Bone marrow mesenchymal stem cells induce angiogenesis and attenuate the remodeling of diabetic cardiomyopathy. Exp Clin Endocrinol Diabetes 116(02):104–111

    CAS  Article  PubMed  Google Scholar 

  14. 14.

    Yang Z, Li K, Yan X, Dong F, Zhao C (2010) Amelioration of diabetic retinopathy by engrafted human adipose-derived mesenchymal stem cells in streptozotocin diabetic rats. Graefe’s Arch Clin Exp Ophthalmol 248(10):1415–1422

    Article  Google Scholar 

  15. 15.

    Shibata T, Naruse K, Kamiya H, Kozakae M, Kondo M, Yasuda Y, Nakamura N, Ota K, Tosaki T, Matsuki T (2008) Transplantation of bone marrow-derived mesenchymal stem cells improves diabetic polyneuropathy in rats. Diabetes 57(11):3099–3107

    PubMed Central  CAS  Article  PubMed  Google Scholar 

  16. 16.

    Ezquer FE, Ezquer ME, Parrau DB, Carpio D, Yañez AJ, Conget PA (2008) Systemic administration of multipotent mesenchymal stromal cells reverts hyperglycemia and prevents nephropathy in type 1 diabetic mice. Biol Blood Marrow Transpl 14(6):631–640

    CAS  Article  Google Scholar 

  17. 17.

    Wu Y, Chen L, Scott PG, Tredget EE (2007) Mesenchymal stem cells enhance wound healing through differentiation and angiogenesis. Stem Cells 25(10):2648–2659

    CAS  Article  PubMed  Google Scholar 

  18. 18.

    Wells JM, Melton DA (2000) Early mouse endoderm is patterned by soluble factors from adjacent germ layers. Development 127(8):1563–1572

    CAS  PubMed  Google Scholar 

  19. 19.

    Ripps H, Shen W (2012) Review: taurine: a “very essential” amino acid. Mol Vis 18:2673

    PubMed Central  CAS  PubMed  Google Scholar 

  20. 20.

    Sturman JA, Hayes KC (1980) The biology of taurine in nutrition and development. Adv Nutr Res 3:231–299

    CAS  Article  Google Scholar 

  21. 21.

    Udawatte C, Qian H, Mangini NJ, Kennedy BG, Ripps H (2008) Taurine suppresses the spread of cell death in electrically coupled RPE cells. Mol Vis 14:1940

    PubMed Central  CAS  PubMed  Google Scholar 

  22. 22.

    Kulakowski EC, Maturo J (1984) Hypoglycemic properties of taurine: not mediated by enhanced insulin release. Biochem Pharmacol 33(18):2835–2838

    CAS  Article  PubMed  Google Scholar 

  23. 23.

    Demeterco C, Beattie GM, Dib SA, Lopez AD, Hayek A (2000) A role for activin A and betacellulin in human fetal pancreatic cell differentiation and growth. J Clin Endocrinol Metab 85(10):3892–3897

    CAS  PubMed  Google Scholar 

  24. 24.

    Kroon E, Martinson LA, Kadoya K, Bang AG, Kelly OG, Eliazer S, Young H, Richardson M, Smart NG, Cunningham J (2008) Pancreatic endoderm derived from human embryonic stem cells generates glucose-responsive insulin-secreting cells in vivo. Nat Biotechnol 26(4):443–452

    CAS  Article  PubMed  Google Scholar 

  25. 25.

    D’Amour KA, Bang AG, Eliazer S, Kelly OG, Agulnick AD, Smart NG, Moorman MA, Kroon E, Carpenter MK, Baetge EE (2006) Production of pancreatic hormone-expressing endocrine cells from human embryonic stem cells. Nat Biotechnol 24(11):1392–1401

    Article  PubMed  Google Scholar 

  26. 26.

    Goicoa S, Álvarez S, Ricordi C, Inverardi L, Domínguez-Bendala J (2006) Sodium butyrate activates genes of early pancreatic development in embryonic stem cells. Cloning Stem Cells 8(3):140–149

    CAS  Article  PubMed  Google Scholar 

  27. 27.

    Bhandari DR, Seo K-W, Sun B, Seo M-S, Kim H-S, Seo Y-J, Marcin J, Forraz N, Roy HL, Larry D (2011) The simplest method for in vitro β-cell production from human adult stem cells. Differentiation 82(3):144–152

    CAS  Article  PubMed  Google Scholar 

  28. 28.

    Ebert R, Ulmer M, Zeck S, Meissner-Weigl J, Schneider D, Stopper H, Schupp N, Kassem M, Jakob F (2006) Selenium supplementation restores the antioxidative capacity and prevents cell damage in bone marrow stromal cells in vitro. Stem Cells 24(5):1226–1235

    CAS  Article  PubMed  Google Scholar 

  29. 29.

    Kolb H, Burkart V (1999) Nicotinamide in type 1 diabetes. Mechanism of action revisited. Diabetes Care 22:B16

    PubMed  Google Scholar 

  30. 30.

    Soria B (2001) In-vitro differentiation of pancreatic β-cells. Differentiation 68(4):205–219

    CAS  Article  PubMed  Google Scholar 

  31. 31.

    Sun Y, Chen L, X-g H, W-k H, J-j D, Sun L, K-x T, Wang B, Song J, Li H (2007) Differentiation of bone marrow-derived mesenchymal stem cells from diabetic patients into insulin-producing cells in vitro. Chin Med J 120(9):771

    CAS  PubMed  Google Scholar 

  32. 32.

    Xu G, Stoffers DA, Habener JF, Bonner-Weir S (1999) Exendin-4 stimulates both beta-cell replication and neogenesis, resulting in increased beta-cell mass and improved glucose tolerance in diabetic rats. Diabetes 48(12):2270–2276

    CAS  Article  PubMed  Google Scholar 

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The authors would like to thank the Iranian National Science Foundation (INSF) for supporting this project under contract no: 90006892.

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Correspondence to Mohammad Taghikhani or Masoud Soleimani.

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Jafarian, A., Taghikhani, M., Abroun, S. et al. Generation of high-yield insulin producing cells from human bone marrow mesenchymal stem cells. Mol Biol Rep 41, 4783–4794 (2014).

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  • Diabetes mellitus
  • Mesenchymal stem cells
  • Differentiation
  • Insulin producing cells
  • Transcription factors