Abstract
Bone marrow-derived mesenchymal stem cells (BMSCs) have the ability to differentiate into insulin-producing cells (IPCs). Bio-scaffolds derived from decellularized organs can act as a carrier for seed cells and may have broad applications in regenerative medicine. This study investigated the effect of native pancreatic stroma obtained from decellularized pancreas on the proliferation, migration and differentiation of BMSCs into IPCs, and explored the potential underlying molecular mechanism. The decellularized pancreas bio-scaffold was obtained by perfusion with Triton X-100/ammonium hydroxide, followed by digestion with a mixture of pepsin and hydrochloric acid to prepare the stroma solution. Islet-like cells were differentiated from BMSCs by a three-step induction method. The differences on the cytological behavior with or without stroma were evaluated by morphological observation, insulin release assay, qRT-PCR assay and western blot analysis. Our results showed that, stroma derived from decellularized pancreas could promote the proliferation and migration of BMSCs. Furthermore, the formation of IPCs could also be promoted, which possessed similar morphology to endogenous islets. During the induced differentiation process, the presence of stroma significantly increased the expression of insulin 1, insulin 2 and Pdx-1, as well as insulin release. This was accompanied by an increase in the phosphorylation of Akt and ERK in third stage cell clusters, which was prevented by the addition of the inhibitors PD98059 and LY294002, respectively. In summary, decellularized pancreatic stroma could promote the proliferation, migration and differentiation of BMSCs into IPCs, and this involved the activation of Akt and ERK signal pathways.
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References
An SY, Han J, Lim HJ, Park SY, Kim JH, Do BR, Kim JH (2014) Valproic acid promotes differentiation of hepatocyte-like cells from whole human umbilical cord-derived mesenchymal stem cells. Tissue Cell 46:127–135. https://doi.org/10.1016/j.tice.2013.12.006
Baca I, Feurle GE, Klempa I, Ziegler A, Schusdziarra V (1990) Morphometry and function of islet cells after different forms of drainage at pancreatic transplantation in rats. Eur Surg Res 22:151–159
Beattie GM et al (2002) A novel approach to increase human islet cell mass while preserving beta-cell function. Diabetes 51:3435–3439
Chang CF, Hsu KH, Chiou SH, Ho LL, Fu YS, Hung SC (2008) Fibronectin and pellet suspension culture promote differentiation of human mesenchymal stem cells into insulin producing cells. J Biomed Mater Res Part A 86:1097–1105. https://doi.org/10.1002/jbm.a.31767
Consolo F et al (2016) A dynamic distention protocol for whole-organ bladder decellularization: histological and biomechanical characterization of the acellular matrix. J Tissue Eng Regen Med 10:E101–E112. https://doi.org/10.1002/term.1767
D’Amour KA et al (2006) Production of pancreatic hormone-expressing endocrine cells from human embryonic stem cells. Nat Biotechnol 24:1392–1401. https://doi.org/10.1038/nbt1259
Davis NE, Beenken-Rothkopf LN, Mirsoian A, Kojic N, Kaplan DL, Barron AE, Fontaine MJ (2012) Enhanced function of pancreatic islets co-encapsulated with ECM proteins and mesenchymal stromal cells in a silk hydrogel. Biomaterials 33:6691–6697. https://doi.org/10.1016/j.biomaterials.2012.06.015
Dickinson RB, Guido S, Tranquillo RT (1994) Biased cell migration of fibroblasts exhibiting contact guidance in oriented collagen gels. Ann Biomed Eng 22:342–356
Digiacomo G, Tusa I, Bacci M, Cipolleschi MG, Dello Sbarba P, Rovida E (2016) Fibronectin induces macrophage migration through a SFK-FAK/CSF-1R pathway. Cell Adhes Migr. https://doi.org/10.1080/19336918.2016.1221566
El-Naggar MM, Al-Mashat FM, Elayat AA, Sibiany AR, Ardawi MS, Badawoud MH (2006) Effect of thawing rate and post-thaw culture on the cryopreserved fetal rat islets: functional and morphological correlation. Life Sci 78:1925–1932. https://doi.org/10.1016/j.lfs.2005.08.043
Goh SK et al (2013) Perfusion-decellularized pancreas as a natural 3D scaffold for pancreatic tissue and whole organ engineering. Biomaterials 34:6760–6772. https://doi.org/10.1016/j.biomaterials.2013.05.066
Hanada M, Feng J, Hemmings BA (2004) Structure, regulation and function of PKB/AKT–a major therapeutic target. Biochem Biophys Acta 1697:3–16. https://doi.org/10.1016/j.bbapap.2003.11.009
Johnson GL, Lapadat R (2002) Mitogen-activated protein kinase pathways mediated by ERK, JNK, and p38 protein kinases. Science 298:1911–1912. https://doi.org/10.1126/science.1072682
Lin HY, Tsai CC, Chen LL, Chiou SH, Wang YJ, Hung SC (2010) Fibronectin and laminin promote differentiation of human mesenchymal stem cells into insulin producing cells through activating Akt and ERK. J Biomed Sci 17:56. https://doi.org/10.1186/1423-0127-17-56
Loneker AE, Faulk DM, Hussey GS, D’Amore A, Badylak SF (2016) Solubilized liver extracellular matrix maintains primary rat hepatocyte phenotype in-vitro. J Biomed Mater Res Part A 104:1846–1847. https://doi.org/10.1002/jbm.a.35778
Nagata N et al (2001) Evaluation of insulin secretion of isolated rat islets cultured in extracellular matrix. Cell Transplant 10:447–451
Navarro-Alvarez N et al (2008) Reestablishment of microenvironment is necessary to maintain in vitro and in vivo human islet function. Cell Transplant 17:111–119
Park KM et al (2016) Decellularized liver extracellular matrix as promising tools for transplantable bioengineered liver promotes hepatic lineage commitments of induced pluripotent stem cells. Tissue Eng Part A 22:449–460. https://doi.org/10.1089/ten.TEA.2015.0313
Petersen TH et al (2010) Tissue-engineered lungs for in vivo implantation. Science 329:538–541. https://doi.org/10.1126/science.1189345
Rao Pattabhi S, Martinez JS, Keller TC 3rd (2014) Decellularized ECM effects on human mesenchymal stem cell stemness and differentiation. Differentiation 88:131–143. https://doi.org/10.1016/j.diff.2014.12.005
Rowlands AS, George PA, Cooper-White JJ (2008) Directing osteogenic and myogenic differentiation of MSCs: interplay of stiffness and adhesive ligand presentation. Am J Physiol Cell Physiol 295:C1037–C1044. https://doi.org/10.1152/ajpcell.67.2008
Shahjalal HM et al (2014) Generation of insulin-producing beta-like cells from human iPS cells in a defined and completely xeno-free culture system. J Mol Cell Biol 6:394–408. https://doi.org/10.1093/jmcb/mju029
Shapiro AM et al (2000) Islet transplantation in seven patients with type 1 diabetes mellitus using a glucocorticoid-free immunosuppressive regimen. N Engl J Med 343:230–238. https://doi.org/10.1056/NEJM200007273430401
Shupe T, Williams M, Brown A, Willenberg B, Petersen BE (2010) Method for the decellularization of intact rat liver. Organogenesis 6:134–136
Smink AM, Faas MM, de Vos P (2013) Toward engineering a novel transplantation site for human pancreatic islets. Diabetes 62:1357–1364. https://doi.org/10.2337/db12-1553
Stendahl JC, Kaufman DB, Stupp SI (2009) Extracellular matrix in pancreatic islets: relevance to scaffold design and transplantation. Cell Transplant 18:1–12
Tang DQ, Cao LZ, Burkhardt BR, Xia CQ, Litherland SA, Atkinson MA, Yang LJ (2004) In vivo and in vitro characterization of insulin-producing cells obtained from murine bone marrow. Diabetes 53:1721–1732
van Dijk A, Niessen HW, Ursem W, Twisk JW, Visser FC, van Milligen FJ (2008) Accumulation of fibronectin in the heart after myocardial infarction: a putative stimulator of adhesion and proliferation of adipose-derived stem cells. Cell Tissue Res 332:289–298. https://doi.org/10.1007/s00441-008-0573-0
Wainwright JM, Czajka CA, Patel UB, Freytes DO, Tobita K, Gilbert TW, Badylak SF (2010) Preparation of cardiac extracellular matrix from an intact porcine heart. Tissue Eng Part C Methods 16:525–532. https://doi.org/10.1089/ten.TEC.2009.0392
Wang RN, Rosenberg L (1999) Maintenance of beta-cell function and survival following islet isolation requires re-establishment of the islet-matrix relationship. J Endocrinol 163:181–190
Williams CM, Engler AJ, Slone RD, Galante LL, Schwarzbauer JE (2008) Fibronectin expression modulates mammary epithelial cell proliferation during acinar differentiation. Can Res 68:3185–3192. https://doi.org/10.1158/0008-5472.CAN-07-2673
Wu D et al (2015) 3D culture of MIN-6 cells on decellularized pancreatic scaffold: vitro and in vivo study. BioMed Res Int 2015:432645. https://doi.org/10.1155/2015/432645
Xu T et al (2015) Three-dimensional culture of mouse pancreatic islet on a liver-derived perfusion-decellularized bioscaffold for potential clinical application. J Biomater Appl 30:379–387. https://doi.org/10.1177/0885328215587610
Zhang Y, Dou Z (2014) Under a nonadherent state, bone marrow mesenchymal stem cells can be efficiently induced into functional islet-like cell clusters to normalize hyperglycemia in mice: a control study. Stem Cell Res Ther 5:66. https://doi.org/10.1186/scrt455
Zhou P et al (2016) The dynamic three-dimensional culture of islet-like clusters in decellularized liver scaffolds. Cell Tissue Res 365:157–171. https://doi.org/10.1007/s00441-015-2356-8
Acknowledgements
This research was supported by grants from National Natural Science Foundation of China (Nos. 81471801, 81672903, 81502569), Medical Innovation Team Program of Jiangsu Province, Science and technology project of Nantong City (MS12018077).
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Guo, Y., Chen, S., Xu, L. et al. Decellularized and solubilized pancreatic stroma promotes the in vitro proliferation, migration and differentiation of BMSCs into IPCs. Cell Tissue Bank 20, 389–401 (2019). https://doi.org/10.1007/s10561-019-09777-4
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DOI: https://doi.org/10.1007/s10561-019-09777-4