Skip to main content

Advertisement

SpringerLink
Log in

Three-dimensional differentiation of adipose-derived mesenchymal stem cells into insulin-producing cells

  • Regular Article
  • Published:
Cell and Tissue Research Aims and scope Submit manuscript

Abstract

The aim of this study is to evaluate the collagen/hyaluronic acid (Col/HA) scaffold effect on the differentiation of insulin-producing cells (IPCs) from adipose-derived mesenchymal stem cells (ASCs). In this experimental study, ASCs were cultured and seeded in a Col/HA scaffold (3D culture) and then treated with induction media. After induction, the presence of IPCs was evaluated using gene expression (PDX-1, GLUT-2 and insulin) analysis and immunocytochemistry, while functional maturity was determined by measuring insulin release in response to low- and high-glucose media. The induced IPCs were morphologically similar to pancreatic islet-like cells. Expression of the islet-associated genes PDX-1, GLUT-2 and insulin genes in 3D-cultured cells was markedly higher than the 2D-cultured cells exposure differentiation media. Compared to the 2D culture of ASCs-derived IPCs, the insulin release from 3D ASCs-derived IPCs showed a nearly 4-fold (p < 0.05) increase when exposed to a high glucose (25 mmol) medium. The percentage of insulin-positive cells in the 3D experimental group showed an approximately 4-fold increase compared to the 2D experimental culture cells. The results of this study demonstrated that the COL/HA scaffold can enhance the differentiation of IPCs from rat ASCs.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Aloysious N, Nair PD (2014) Enhanced survival and function of islet-like clusters differentiated from adipose stem cells on a three-dimensional natural polymeric scaffold: an in vitro study. Tissue Eng Part A 20:1508–22

    Article  CAS  PubMed  Google Scholar 

  • Bayati V, Hashemitabar M, Gazor R, Nejatbakhshi R, Bijannejad D (2013) Expression of surface markers and muogenic potential of rat bone marrow-and adipose derived stem cell a comparative study. Anat Cell Biol 46:113–121

    Article  PubMed Central  PubMed  Google Scholar 

  • Bonner-Weir S, Taneja M, Weir GC, Tatarkiewicz K, Song KH, Sharma A, O’Neil JJ (2000) In vitro cultivation of human islets from expanded ductal tissue. Proc Natl Acad Sci U S A 97:7999–8004

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Bose B, Shenoy SP, Konda S, Wangikar P (2012) Human embryonic stem cell differentiation into insulin secreting β-cells for diabetes. Cell Biol Int 36:1013–20

    Article  CAS  PubMed  Google Scholar 

  • Brun T, Franklin I, St-Onge L, Biason-Lauber A, Schoenle EJ, Wollheim CB, Gauthier BR (2004) The diabetes-linked transcription factor PAX4 promotes b-cell proliferation and survival in rat and human islets. J Cell Biol 167:1123–1135

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Calderon L, Collin E, Velasco-Bayon D, Murphy M, O’Halloran D, Pandit A (2010) Type II collagen-hyaluronan hydrogel-a step towards a scaffold for intervertebral disc tissue engineering. Eur Cell Mater 20:134–48

    CAS  PubMed  Google Scholar 

  • Chen PY, Huang LL, Hsieh HJ (2007) Hyaluronan preserves the proliferation and differentiation potentials of long-term cultured murine adipose-derived stromal cells. Biochem Biophys Res Commun 360:1–6

    Article  CAS  PubMed  Google Scholar 

  • Chun SY, Mack DL, Moorefield E, Oh SH, Kwon TG, Pettenati MJ, Yoo JJ, Coppi PD, Atala A, Soker S (2012) Pdx1 and controlled culture conditions induced differentiation of human amniotic fluid-derived stem cells to insulin-producing clusters. J Tissue Eng Regen Med. doi:10.1002/term.1631

    PubMed Central  Google Scholar 

  • Cras-Méneur C, Elghazi L, Czernichow P, Scharfmann R (2001) Epidermal growth factor increases undifferentiated pancreatic embryonic cells in vitro: a balance between proliferation and differentiation. Diabetes 50:1571–9

    Article  PubMed  Google Scholar 

  • Davidenko N, Campbell JJ, Thian ES, Watson CJ, Cameron RE (2010) Collagen-hyaluronic acid scaffolds for adipose tissue engineering. Acta Biomater 6:3957–68

    Article  CAS  PubMed  Google Scholar 

  • Dunbar AJ, Goddard C (2000) Structure-function and biological role of betacellulin. Int J Biochem Cell Biol 32:805–15

    Article  CAS  PubMed  Google Scholar 

  • Ebrahimie M, Esmaeili F, Cheraghi C, Houshmand F, Shabani L, Ebrahimie E (2014) Efficient and simple production of insulin-producing cells from embryonal carcinoma stem cells using mouse neonate pancreas extract as a natural inducer. PLoS ONE 9:e90885

    Article  PubMed Central  PubMed  Google Scholar 

  • Edlund H (1998) Transcribing pancreas. Diabetes 47:1817–1823

    Article  CAS  PubMed  Google Scholar 

  • Eyre DR, Matsui Y, Wu JJ (2002) Collagen polymorphisms of the intervertebral disc. Biochem Soc Trans 30:844–848

    Article  CAS  PubMed  Google Scholar 

  • Florio P, Luisi S, Marchetti P, Lupi R, Cobellis L, Falaschi C, Sugino H, Navalesi R, Genazzani AR, Petraglia F (2000) Activin A stimulates insulin secretion in cultured human pancreatic islets. J Endocrinol Investig 23:231–234

    Article  CAS  Google Scholar 

  • Gabr MM, El-Far MA, Ibrahim RY, El-Halawani SM, Ismail AM, Zakaria MM, Sherry MK, Ghoneim MA (2012) Biochemical induction of adult bone marrow hematopeitic stem cells into insulin producing cells. Curr Top Biochem Res 14:45–56

    CAS  Google Scholar 

  • Grayson WL, Ma T, Bunnell B (2004) Human mesenchymal stem cells tissue development in 3D PET matrices. Biotechnol Prog 20:905–12

    Article  CAS  PubMed  Google Scholar 

  • Guo W, Miao C, Liu S, Qiu Z, Li J, Duan E (2009) Efficient differentiation of insulin-producing cells from skin-derived stem cells. Cell Prolif 42:49–62

    Article  CAS  PubMed  Google Scholar 

  • Hammar EB, Irminger JC, Rickenbach K, Parnaud G, Ribaux P, Bosco D, Rouiller DG, Halban PA (2005) Activation of NF-kB by extracellular matrix is involved in spreading and glucose-stimulated insulin secretion of pancreatic b cells. J Biol Chem 280:30630–30637

    Article  CAS  PubMed  Google Scholar 

  • Hardikar AA, Marcus-Samuels B, Geras-Raaka E, Raaka BM, Gershengorn MC (2003) Human pancreatic precursor cells secrete FGF2 to stimulate clustering into hormone expressing islet-like cell aggregates. Proc Natl Acad Sci USA 100:7117–7122

  • Herbst RS (2004) Review of epidermal growth factor receptor biology. Int J Radiat Oncol Biol Phys 59:21–6

    Article  CAS  PubMed  Google Scholar 

  • Huotari MA, Palgi J, Otonkoski T (1998) Growth factor-mediated proliferation and di€erentiation of insulin-pro- ducing INS-1 and RINm5F cells; identification of betacellulin as a novel b-cell mitogen. Endocrinology 139:1494–1499

    CAS  PubMed  Google Scholar 

  • Kayoko TO, Yoshinori O, Maki M, Akio H, Akira A, Akira T, Abe K, Tamaki T (2010) Micro 3D culture system using hyaluronan-collagen capsule for skeletal muscle-derived stem cells. Open Tissue Eng Regen Med J 3:18–27

    Article  Google Scholar 

  • Khorsandi L, Mirhoseini M, Mohamadpour M, Orazizadeh M, Khaghani S (2013) Effect of curcumin on dexamethasone-induced testicular toxicity in mice. Pharm Biol 51:206–12

    Article  CAS  PubMed  Google Scholar 

  • Knight KR, Uda Y, Findlay MW, Brown DL, Cronin KJ, Jamieson E, Tai T, Keramidaris E, Penington AJ, Rophael J, Harrison LC, Morrison WA (2006) Vascularized tissue-engineered chambers promote survival and function of transplanted islets and improve glycemic control. FASEB J 20:565

    CAS  PubMed  Google Scholar 

  • Liu SH, Lee LT (2012) Efficient differentiation of mouse embryonic stem cells into insulin-producing cells. Exp Diabetes Res 2012:201295

    PubMed Central  PubMed  Google Scholar 

  • Liu J, Liu Y, Wang H, Hao H, Han Q, Shen J, Shi J, Li C, Mu Y, Han W (2013) Direct differentiation of hepatic stem-like WB cells into insulin-producing cells using small molecules. Sci Rep 3:1–8

    CAS  Google Scholar 

  • Lumelsky N, Blondel O, Laeng P, Velasco I, Ravin R, McKay R (2001) Differentiation of embryonic stem cells to insulin-secreting structures similar to pancreatic islets. Science 292:1389–1394

    Article  CAS  PubMed  Google Scholar 

  • Maleki A, Kjoniksen AL, Nystrom B (2007) Characterization of the chemical degradation of hyaluronic acid during chemical gelation in the presence of different cross-linker agents. Carbohydr Res 342:2776–2792

    Article  CAS  PubMed  Google Scholar 

  • Mashima H, Shibata H, Mine T, Kojima I (1996) Formation of insulin-producing cells from pancreatic acinar AR42J cells by hepatocyte growth factor. Endocrinology 137:3969–76

    CAS  PubMed  Google Scholar 

  • Mason MN, Arnold CA, Mahoney MJ (2009) Entrapped collagen type 1 promotes differentiation of embryonic pancreatic precursor cells into glucose-responsive beta-cells when cultured in three-dimensional PEG hydrogels. Tissue Eng Part A 15:3799–808

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Mohr JC, de Pablo JJ, Palecek SP (2006) 3-D microwell culture of human embryonic stem cells. Biomaterials 27:6032–42

    Article  CAS  PubMed  Google Scholar 

  • Mukherjee A, Sidis Y, Mahan A, Raher MJ, Xia Y, Rosen ED, Bloch KD, Thomas MK, Schneyer AL (2007) FSTL3 deletion reveals roles for TGFbeta family ligands in glucose and fat homeostasis in adults. Proc Natl Acad Sci U S A 104:1348–1353

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Olson AL, Pessin JE (1996) Structure function, and regulation of the mammalian facilitative glucose transporter gene family. Annu Rev Nutr 16:235–56

    Article  CAS  PubMed  Google Scholar 

  • Pandey S (2010) Stem cell transplantation: a future for diabetic paitient. Int J Pharm Sci Rev Res 1:68–71

    Google Scholar 

  • Park MK, Han C, Lee KH, Hong SH, Kim HS, Lee YJ, Jeong IK, Noh JH, Yang TY, Lee MS, Kim KW, Lee MK (2007) Effects of activin A on pancreatic ductal cells in streptozotocin-induced diabetic rats. Transplantation 83:925–930

    Article  CAS  PubMed  Google Scholar 

  • Parnaud G, Hammar E, Ribaux P, Donath MY, Berney T, Halban PA (2009) Signaling pathways implicated in the stimulation of b-cell proliferation by extracellular matrix. Mol Endocrinol 23:1264–1271

    Article  CAS  PubMed  Google Scholar 

  • Peshavaria M, Cissell MA, Henderson E, Petersen HV, Stein R (2000) The PDX-1 activation domain provides specific functions necessary for transcriptional stimulation in pancreatic beta-cells. Mol Endocrinol 14:1907–1917

    CAS  PubMed  Google Scholar 

  • Puissant B, Barreau C, Bourin P, Clavel C, Corre J, Bousquet C, Taureau C, Cousin B, Abbal M, Laharrague P, Penicaud L, Casteilla L, Blancher A (2005) Immunomodulatory effect of human adipose tissue-derived adult stem cells: comparison with bone marrow mesenchymal stem cells. Br J Haematol 129:118–29

    Article  PubMed  Google Scholar 

  • Richardson SM, Walker RV, Parker S, Rhodes NP, Hunt JA, Freemont AJ, Hoyland JA (2006) Intervertebral disc cell-mediated mesenchymal stem cell differentiation. Stem Cells 24:707–716

    Article  CAS  PubMed  Google Scholar 

  • Risbud MV, Bhonde RR (2001) Suitability of cellulose molecular dialysis membrane for bioartificial pancreas: in vitro biocompatibility studies. J Biomed Mater Res 54:436–444

    Article  CAS  PubMed  Google Scholar 

  • Rooney P, Kumar S (1993) Inverse relationship between hyaluronan and collagens in development and angiogenesis. Differentiation 54:1–9

    CAS  PubMed  Google Scholar 

  • Schmeichel KL, Bissell MJ (2003) Modeling tissue-specific signaling and organ function in three dimensions. J Cell Sci 116:2377–88

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Skoudy A, Rovira M, Savatier P, Martin F, León-Quinto T, Soria B, Real FX (2004) Transforming growth factor (TGF) beta, fibroblast growth factor (FGF) and retinoid signalling pathways promote pancreatic exocrine gene expression in mouse embryonic stem cells. Biochem J 379:749–56

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Speier S, Gjinovci A, Charollais A, Meda P, Rupnik M (2007) Cx36-mediated coupling reduces b-cell heterogeneity, confines the stimulating glucose concentration range, and affects insulin release kinetics. Diabetes 56:1078–1086

    Article  CAS  PubMed  Google Scholar 

  • Sun Y, Chen L, Hou XG, Hou WK, Dong JJ, Sun L, Tang KX, Wang B, Song J, Li H, Wang KX (2007) Differentiation of bone marrow-derived mesenchymal stem cells from diabetic patients into insulin-producing cells in vitro. Chin Med J (Engl) 120:771–6

    CAS  Google Scholar 

  • Takeuchi H, Nakatsuji N, Suemori H (2014) Endodermal differentiation of human pluripotent stem cells to insulin-producing cells in 3D culture. Sci Rep 4:4488

    PubMed Central  PubMed  Google Scholar 

  • Tang S, Vickers SM, Hsu HP, Spector M (2007) Fabrication and characterization of porous hyaluronic acid-collagen composite scaffolds. J Biomed Mater Res A 82:323–335

    Article  PubMed  Google Scholar 

  • Tholpady SS, Katz AJ, Ogle RC (2003) Mesenchymal stem cells from rat visceral fat exhibit multipotential differentiation in vitro. Anat Rec A 272:398–402

    Article  CAS  Google Scholar 

  • Wang X, Ye K (2009) Three-dimensional differentiation of embryonic stem cells into islet-like insulin-producing clusters. Tissue Eng Part A 15:1941–52

    Article  CAS  PubMed  Google Scholar 

  • Wang LX, Zhao H, Jiang B, Ding Y (2009) Adhesion and growth of human periodontal ligament cells on hyaluronic acid/collagen scaffold. Hua Xi Kou Qiang Yi Xue Za Zhi 27:220–3

    PubMed  Google Scholar 

  • Watada H, Kajimoto Y, Miyagawa J, Hanafusa T, Hamaguchi K, Matsuoka T, Yamamoto K, Matsuzawa Y, Kawamori R, Yamasaki Y (1996) PDX-1 induces insulin and glucokinase gene expressions in alphaTC1 clone 6 cells in the presence of betacellulin. Diabetes 45:1826–31

    Article  CAS  PubMed  Google Scholar 

  • Watt FM, Hogan BL (2000) Out of Eden: stem cells and their niches. Science 287:1427–30

    Article  CAS  PubMed  Google Scholar 

  • Weber LM, Hayda KN, Anseth KS (2008) Cell-matrix interactions improve b-cell survival and insulin secretion in three-dimensional culture. Tissue Eng Part A 14:1959–1968

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Wong RS (2011) Extrinsic factors involved in the differentiation of stem cells into insulin-producing cells: an overview. Exp Diabetes Res. doi:10.1155/2011/406182

    PubMed Central  PubMed  Google Scholar 

  • Woods EJ, Walsh CM, Sidner RA, Zieger MA, Lakey JR, Ricordi C, Critser JK (2004) Improved in vitro function of islets using small intestinal submucosa. Transplant Proc 36:1175

    Article  CAS  PubMed  Google Scholar 

  • Xu W, Hu R, Fan E, Han D (2011) Adipose-derived mesenchymal stem cells in collagen-hyaluronic acid gel composite scaffolds for vocal fold regeneration. Ann Otol Rhinol Laryngol 120:123–30

    Article  PubMed  Google Scholar 

  • Yang L, Li S, Hatch H, Ahrens K, Cornelius JG, Petersen BE (2002) In vitro transdifferentiation of adult hepatic stem cells into pancreatic endocrine hormoneproducing cells. Proc Natl Acad Sci U S A 99:8078–8083

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Yang Z, Chen Z, Liu K, Bai Y, Jiang T, Feng D, Feng G (2013) Experimental study on tissue engineered cartilage complex three-dimensional nano-scaffold with collagen type II and hyaluronic acid in vitro. Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi 27:1240–5

    CAS  PubMed  Google Scholar 

  • Zahran F, Salama M, Lotfy A, El-Deen IM (2012) Isolation and characterization of adipose tissue-derived stem cells: an in vitro study. Basic Res J Med Clin Sci 1:88–94

    Google Scholar 

  • Zalzman M, Anker-Kitai L, Efrat S (2005) Differentiation of human liver-derived, insulin-producing cells toward the cell phenotype. Diabetes 54:2568–75

    Article  CAS  PubMed  Google Scholar 

  • Zavan B, Cortivo R, Tonello C, Abatangelo G (2003) Gland cell cultures into 3D hyaluronan-based scaffolds. J Mater Sci Mater Med 14:727–729

    Article  CAS  PubMed  Google Scholar 

  • Zhang H, Wei YT, Tsang KS, Sun CR, Li J, Huang H, Cui FZ, An YH (2008) Implantation of neural stem cells embedded in hyaluronic acid and collagen composite conduit promotes regeneration in a rabbit facial nerve injury model. J Transl Med 6:67

    Article  PubMed Central  PubMed  Google Scholar 

  • Zhao FQ, Keating AF (2007) Functional properties and genomics of glucose transporters. Curr Genomics 8:113–128

    Article  PubMed Central  CAS  PubMed  Google Scholar 

Download references

Acknowledgment

The financial cost of this project has been provided by the research council of the Ahvaz Jundishapur University of Medical Sciences (Grant number: CMRC-48).

Author information

Authors and Affiliations

  1. Cell & Molecular Research Center, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran, P.O. Box: 61335

    Layasadat Khorsandi, Ali Khodadadi, Fereshteh Nejad-Dehbashi & Sadegh Saremy

  2. Department of Anatomical Sciences, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran

    Layasadat Khorsandi

  3. Department of Immunology and Petroleum and Environmental Pollutants Research Center, Faculty of Medicine, Jundishapur University of Medical Sciences, Ahvaz, Iran

    Ali Khodadadi

Authors
  1. Layasadat Khorsandi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ali Khodadadi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Fereshteh Nejad-Dehbashi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Sadegh Saremy
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Layasadat Khorsandi.

Electronic supplementary material

Below is the link to the electronic supplementary material.

ESM 1

(XML 9 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Khorsandi, L., Khodadadi, A., Nejad-Dehbashi, F. et al. Three-dimensional differentiation of adipose-derived mesenchymal stem cells into insulin-producing cells. Cell Tissue Res 361, 745–753 (2015). https://doi.org/10.1007/s00441-015-2140-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00441-015-2140-9

Keywords

Search

Navigation