Differentiation Potential of Mouse Embryonic Stem Cells into Insulin Producing Cells in Pancreatic Islet Microenvironment

 

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Abstract

Background: The differentiation capacity of embryonic stem cells (ESCs) has great promise for type-1 diabetes for cellular treatment. Therefore, different strategies have been reported so far for derivation of insulin producing cells (IPCs) from ESCs. Providing similar microenvironmental conditions as in vivo, functional differentiation of stem cells into desired cell types could be obtained in vitro. The aim of the present research was to utilize differentiation potential of ESCs to IPCs by co-culture with mouse pancreatic islets (mPIs) for the first time.

Methods: We present an in-direct differentiation protocol which compared with a conventional differentiation protocol. Novel in-direct co-culture differentiation protocol in which mPIs induced differentiation of ESCs into IPCs was used. This technique was compared with the chemical differentiation protocol that involved supplementing the differentiation media with specific growth factors. We analyzed differentiated cells in both groups by immune labelling, gene expression and protein secretion.

Results: IPCs were obtained with in-direct co-culture within 30 days. Differentiated ESCs were found to be positive for IPC specific markers, Pdx1, Insulin, C-peptide, Glut2 and MafA. The results of immunocytochemical and gene expression analysis showed higher differentiation efficiency in co-culture group than chemical differentiation group. These results were confirmed by the response assay to high glucose levels with ELISA for insulin.

Discussion: Our findings illustrate the significant effect of co-culture in different stages of differentiation and maturation of ESCs in vitro. We have developed an efficient and easy way to differentiate ESCs into IPCs, which possess similar characters of mature insulin positive cells.

• References

• 1 Whiting DR, Guariguata L, Weil C et al. IDF diabetes atlas: Global estimates of the prevalence of diabetes for 2011 and 2030. Diabetes Res Clin Pract 2011; 3: 311-321
  PubMedGoogle Scholar
• 2 Masharani U, German MS. Pancreatic hormones and diabetes mellitus. Gardner DG, Shoback DM. (eds.) Greenspan’s Basic & Clinical Endocrinology. 9^th ed. New York: McGraw-Hill Medical; 2011. Chapter 17
  Google Scholar
• 3 Bonner-Weir S, Baxter LA, Schuppin GT et al. A second pathway for regeneration of adult exocrine and endocrine pancreas. A possible recapitulation of embryonic development. Diabetes 1993; 42: 1715-1720
  CrossrefPubMedGoogle Scholar
• 4 Muir KR, Lima MJ, Docherty HM et al. Cell therapy for type 1 diabetes. Q J Med 2014; 107: 253-259
  CrossrefPubMedGoogle Scholar
• 5 Wobus AM, Boheler KR. Embryonic stem cells: Prospects for developmental biology and cell therapy. Physiol rev 2005; 85: 635-678
  CrossrefPubMedGoogle Scholar
• 6 Noguchi H. Stem cells for the treatment of diabetes. Endocrine Journal 2007; 54: 7-16
  CrossrefPubMedGoogle Scholar
• 7 Jacqueline V, Schiesser JV, Wells JM. Generation of β cells from human pluripotent stem cells: Are we there yet?. Ann N Y Acad Sci. Issue: The Year in Diabetes and Obesity 2014; 124-137
  PubMedGoogle Scholar
• 8 Abdelalim EM, Bonnefond A, Griscelli AB et al. Pluripotent stem cells as a potential tool for disease modelling and cell therapy in diabetes. Stem Cell Rev 2014; 10: 327-337
  CrossrefPubMedGoogle Scholar
• 9 Calafiore R, Montanucci P, Basta G. Stem cells for pancreatic b-cell replacement in diabetes mellitus: actual perspectives. Curr Opin Organ Transplant 2014; 19: 162-168
  CrossrefPubMedGoogle Scholar
• 10 Assady S, Maor G, Amit M et al. Insulin production by human embryonic stem cells. Diabetes 2001; 50: 1691-1697
  CrossrefPubMedGoogle Scholar
• 11 Amit M, Itskovitz-Eldor J. Derivation and spontaneous differentiation of human embryonic stem cells. J Anat 2002; 200: 225-232
  CrossrefPubMedGoogle Scholar
• 12 Blyszczuk P, Czyz J, Kania G et al. Expression of Pax4 in embryonic stem cells promotes differentiation of nestin-positive progenitor and insulin-producing cells. PNAS 2003; 100: 998-1003
  CrossrefPubMedGoogle Scholar
• 13 Lumelsky N, Blondel O, Laeng P et al. Differentiation of embryonic stem cells to insulin-secreting structures similar to pancreatic islets. Science 2001; 292: 1389-1394
  CrossrefPubMedGoogle Scholar
• 14 Miyazaki S, Yamato E, Miyazaki J. Regulated expression of pdx-1 promotes in vitro differentiation of insulin-producing cells from embryonic stem cells. Diabetes 2004; 53: 1030-1037
  CrossrefPubMedGoogle Scholar
• 15 Kania G, Blyszczuk P, Wobus AM. The generation of insulin-producing cells from embryonic stem cells – a discussion of controversial findings. Int J Dev Biol 2004; 48: 1061-1064
  CrossrefPubMedGoogle Scholar
• 16 Kaitsuka T, Noguchi H, Shiraki N et al. Generation of functional insulin-producing cells from mouse embryonic stem cells through 804G cell-derived extracellular matrix and protein transduction of transcription factors. Stem Cells Trans Med 2014; 3: 114-127
  CrossrefPubMedGoogle Scholar
• 17 Lima MJ, Docherty HM, Chen Y et al. Pancreatic transcription factors containing protein transduction domains drive mouse embryonic stem cells towards endocrine pancreas. PLOS ONE 2012; 7: e36481
  CrossrefPubMedGoogle Scholar
• 18 Xu X, Browning VL, Odorico JS. Activin, BMP and FGF pathways cooperate to promote endoderm and pancreatic lineage cell differentiation from human embryonic stem cells. Mech Dev 2011; 128: 412-427
  CrossrefPubMedGoogle Scholar
• 19 Delisle JC, Martignat L, Dubreil L et al. Pdx-1 or Pdx-1-VP16 protein transduction induces β-cell gene expression in liver-stem WB cells. BMC Res Notes 2009; 2: 3
  CrossrefPubMedGoogle Scholar
• 20 Shi Y, Hou L, Tang F et al. Inducing embryonic stem cells to differentiate into pancreatic beta cells by a novel three-step approach with activin A and all-trans retinoic acid. Stem Cells 2005; 23: 656-662
  CrossrefPubMedGoogle Scholar
• 21 D’Amour KA, Bang AG, Eliazer S et al. Production of pancreatic hormone–expressing endocrine cells from human embryonic stem cells. Nat Biotechnol 2006; 24: 1392-1401
  CrossrefPubMedGoogle Scholar
• 22 Jiang J, Au M, Lu K et al. Generation of insulin-producing islet-like clusters from human embryonic stem cells. Stem Cells 2007; 25: 1940-1953
  CrossrefPubMedGoogle Scholar
• 23 Ku HT, Zhang N, Kubo A et al. Committing embryonic stem cells t to early endocrine pancreas in vitro. Stem Cells 2004; 22: 1205-1217
  CrossrefPubMedGoogle Scholar
• 24 Li H, Lam A, Xu A et al. High dosage of exendin-4 increased early insulin secretion in differentiated beta cells from mouse embryonic stem cells. Acta Pharmacol Sin 2010; 31: 570-577
  CrossrefPubMedGoogle Scholar
• 25 Roche E, Jones J, Arribas MI et al. Role of small bioorganic molecules in stem cell differentiation to insulin-producing cells. Bioorg Med Chem 2006; 14: 6466-6474
  CrossrefPubMedGoogle Scholar
• 26 Brolen GKC, Heins N, Edsbagge J et al. Signals from the embryonic mouse pancreas induce differentiation of human embryonic stem cells into insulin-producing β-cell – like cells. Diabetes 2005; 54: 2867-2874
  CrossrefPubMedGoogle Scholar
• 27 Vaca P, Martin F, Vegara-Meseguer JM et al. Induction of differentiation of embryonic stem cells into insulin-secreting cells by fetal soluble factors. Stem Cells 2006; 24: 258-265
  CrossrefPubMedGoogle Scholar
• 28 Banerjee I, Sharma N, Yarmush M. Impact of co-culture on pancreatic differentiation of embryonic stem cells. J Tissue Eng Regen Med 2011; 5: 313-323
  CrossrefPubMedGoogle Scholar
• 29 Karaoz E, Okcu A, Unal ZS et al. Adipose tissue derived mesenchymal stem cells efficiently differentiate into insulin producing cells in pancreatic islet microenvironment both in vitro and in vivo. Cytotherapy 2013; 15: 557-570
  CrossrefPubMedGoogle Scholar
• 30 Karaoz E, Ayhan S, Okcu A et al. Bone marrow-derived mesenchymal stem cells co-cultured with pancreatic islets display β cell plasticity. J Tissue Eng Regen Med 2011; 6: 491-500
  PubMedGoogle Scholar
• 31 Karaoz E, Genc ZS, Demircan PC et al. Protection of rat pancreatic islet function and viability by coculture with rat bone-marrow derived mesenchymal stem cells. Cell Death Dis 2010; 1: e36
  CrossrefPubMedGoogle Scholar
• 32 Xie H, Wang Y, Zhang H et al. Role of injured pancreatic extract promotes bone marrow-derived mesenchymal stem cells efficiently differentiate into insulin-producing cells. PLOS ONE 2013; 8: e76056
  CrossrefPubMedGoogle Scholar
• 33 Wei R, Yang J, Hou W et al. Insulin-producing cells derived from human embryonic stem cells: comparison of definitive endoderm- and nestin-positive progenitor-based differentiation strategies. PLoS ONE 2013; 8: e72513
  CrossrefPubMedGoogle Scholar
• 34 Felicia W, Pagliuca FW, Melton DA. How to make a functional β-cell. Development 2013; 140: 2472-2483
  CrossrefPubMedGoogle Scholar
• 35 Murry CE, Keller G. Differentiation of embryonic stem cells to clinically relevant populations: lessons from embryonic development. Cell 2008; 132: 661-680
  CrossrefPubMedGoogle Scholar
• 36 Josue MK, Luc B. Milestones of pancreatic beta cell differentiation from embryonic stem cells. Adv Gene Mol Cell Ther 2007; 2: 161-171
  PubMedGoogle Scholar
• 37 Shen J, Cheng Y, Han Q et al. Generating insulin-producing cells for diabetic therapy: Existing strategies and new development. Ageing Res Rev 2013; 469-478
  PubMedGoogle Scholar
• 38 Sander M, German MS. The β cell transcription factors and development of the pancreas. J Mol Med 1997; 75: 327-340
  CrossrefPubMedGoogle Scholar
• 39 Bouwens L, Houbracken I, Mfopou JK. The use of stem cells for pancreatic regeneration in diabetes mellitus. Nat Rev Endocrinol 2013; 9: 598-606
  CrossrefPubMedGoogle Scholar
• 40 Ding L, Gysemans C, Mathieu C. β-Cell differentiation and regeneration in type 1 diabetes. Diabetes, Obes Metab 2013; 15: 98-104
  CrossrefPubMedGoogle Scholar
• 41 Stanekzai J, Isenovic ER, Mousa SA. Treatment options for diabetes: Potential role of stem cells. Diabetes Res Clin Pract 2012; 98: 361-368
  CrossrefPubMedGoogle Scholar
• 42 Hang Y, Stein R. MafA and MafB activity in pancreatic β cells. Trends Endocrinol Metab 2011; 22: 364-373
  CrossrefPubMedGoogle Scholar
• 43 Nishimura W, Bonner-Weir S, Sharma A. Expression of MafA in pancreatic progenitors is detrimental for pancreatic development. Dev Biol 2009; 333: 108-120
  CrossrefPubMedGoogle Scholar
• 44 Artner I, Hang Y, Mazur M et al. MafA and MafB Regulate Genes Critical to β-Cells in a Unique Temporal Manner. Diabetes 2010; 59: 2530-2539
  CrossrefPubMedGoogle Scholar

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