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Exp Clin Endocrinol Diabetes 2016; 124(02): 120-129
DOI: 10.1055/s-0035-1554720
Article
© Georg Thieme Verlag KG Stuttgart · New York

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

I. Yilmaz
1   Center for Stem Cell and Gene Therapies Research and Practice, Institute of Health Sciences, Stem Cell Department, Kocaeli University, Kocaeli, Turkey
2   Center for Regenerative Medicine and Stem Cell Research & Manufacturing (LivMedCell), Liv Hospital, Istanbul, Turkey
,
A. E. Sariboyaci
1   Center for Stem Cell and Gene Therapies Research and Practice, Institute of Health Sciences, Stem Cell Department, Kocaeli University, Kocaeli, Turkey
3   Vocational School of Health Services, Eskisehir Osmangazi University, Eskisehir, Turkey
,
C. Subasi
1   Center for Stem Cell and Gene Therapies Research and Practice, Institute of Health Sciences, Stem Cell Department, Kocaeli University, Kocaeli, Turkey
2   Center for Regenerative Medicine and Stem Cell Research & Manufacturing (LivMedCell), Liv Hospital, Istanbul, Turkey
,
E. Karaoz
1   Center for Stem Cell and Gene Therapies Research and Practice, Institute of Health Sciences, Stem Cell Department, Kocaeli University, Kocaeli, Turkey
2   Center for Regenerative Medicine and Stem Cell Research & Manufacturing (LivMedCell), Liv Hospital, Istanbul, Turkey
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Publikationsverlauf

received 24. Dezember 2014
first decision 30. April 2015

accepted 03. Juni 2015

Publikationsdatum:
20. November 2015 (online)

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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.

Key words

differentiation - co-culture - microenvironment - mouse pancreatic islets - embryonic stem cells - insulin producing cells

Supplementary Material

 

  • 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
    PubMedSuche in Google Scholar
  • 2 Masharani U, German MS. Pancreatic hormones and diabetes mellitus. Gardner DG, Shoback DM. (eds.) Greenspan’s Basic & Clinical Endocrinology. 9th ed. New York: McGraw-Hill Medical; 2011. Chapter 17
    Suche in 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
    CrossrefPubMedSuche in Google Scholar
  • 4 Muir KR, Lima MJ, Docherty HM et al. Cell therapy for type 1 diabetes. Q J Med 2014; 107: 253-259
    CrossrefPubMedSuche in Google Scholar
  • 5 Wobus AM, Boheler KR. Embryonic stem cells: Prospects for developmental biology and cell therapy. Physiol rev 2005; 85: 635-678
    CrossrefPubMedSuche in Google Scholar
  • 6 Noguchi H. Stem cells for the treatment of diabetes. Endocrine Journal 2007; 54: 7-16
    CrossrefPubMedSuche in Google 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
    PubMedSuche in Google 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
    CrossrefPubMedSuche in Google 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
    CrossrefPubMedSuche in Google Scholar
  • 10 Assady S, Maor G, Amit M et al. Insulin production by human embryonic stem cells. Diabetes 2001; 50: 1691-1697
    CrossrefPubMedSuche in Google Scholar
  • 11 Amit M, Itskovitz-Eldor J. Derivation and spontaneous differentiation of human embryonic stem cells. J Anat 2002; 200: 225-232
    CrossrefPubMedSuche in Google 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
    CrossrefPubMedSuche in Google 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
    CrossrefPubMedSuche in Google 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
    CrossrefPubMedSuche in Google 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
    CrossrefPubMedSuche in Google 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
    CrossrefPubMedSuche in Google 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
    CrossrefPubMedSuche in Google 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
    CrossrefPubMedSuche in Google 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
    CrossrefPubMedSuche in Google 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
    CrossrefPubMedSuche in Google 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
    CrossrefPubMedSuche in Google 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
    CrossrefPubMedSuche in Google 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
    CrossrefPubMedSuche in Google 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
    CrossrefPubMedSuche in Google 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
    CrossrefPubMedSuche in Google 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
    CrossrefPubMedSuche in Google 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
    CrossrefPubMedSuche in Google 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
    CrossrefPubMedSuche in Google 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
    CrossrefPubMedSuche in Google 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
    PubMedSuche in Google 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
    CrossrefPubMedSuche in Google 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
    CrossrefPubMedSuche in Google 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
    CrossrefPubMedSuche in Google Scholar
  • 34 Felicia W, Pagliuca FW, Melton DA. How to make a functional β-cell. Development 2013; 140: 2472-2483
    CrossrefPubMedSuche in Google Scholar
  • 35 Murry CE, Keller G. Differentiation of embryonic stem cells to clinically relevant populations: lessons from embryonic development. Cell 2008; 132: 661-680
    CrossrefPubMedSuche in Google 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
    PubMedSuche in Google 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
    PubMedSuche in Google Scholar
  • 38 Sander M, German MS. The β cell transcription factors and development of the pancreas. J Mol Med 1997; 75: 327-340
    CrossrefPubMedSuche in Google 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
    CrossrefPubMedSuche in Google Scholar
  • 40 Ding L, Gysemans C, Mathieu C. β-Cell differentiation and regeneration in type 1 diabetes. Diabetes, Obes Metab 2013; 15: 98-104
    CrossrefPubMedSuche in Google 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
    CrossrefPubMedSuche in Google Scholar
  • 42 Hang Y, Stein R. MafA and MafB activity in pancreatic β cells. Trends Endocrinol Metab 2011; 22: 364-373
    CrossrefPubMedSuche in Google 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
    CrossrefPubMedSuche in Google 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
    CrossrefPubMedSuche in Google Scholar