Differentiation of Alginate-Encapsulated Wharton Jelly-Derived Mesenchymal Stem Cells into Insulin Producing Cells

Document Type : Original Article


1 Biology Department, Science and Research Branch, Islamic Azad University, Tehran, Iran

2 Department of Hematology, Faculty of Medical Sciences, Tarbiat Modarres University, Tehran, Iran


Objective: Insulin insufficiency due to the reduced pancreatic beta cell number is the hallmark of diabetes, resulting in
an intense focus on the development of beta-cell replacement options. One approach to overcome the problem is to
search for alternative sources to induce insulin-producing cells (IPCs), the advent of mesenchymal stem cells (MSCs)
holds great promise for producing ample IPCs. Encapsulate the MSCs with alginate improved anti-inflammatory effects
of MSC treatment. This study aimed to evaluate the differentiation of wharton jelly-derived MScs into insulin producing
cells using alginate encapsulation.
Materials and Methods: In this experimental study, we established an efficient IPCs differentiation strategy of human
MSCs derived from the umbilical cord’s Wharton jelly with lentiviral transduction of Pancreas/duodenum homeobox
protein 1 (PDX1) in a 21-day period using alginate encapsulation by poly-L-lysine (PLL) and poly-L-ornithine (PLO)
outer layer. During differentiation, the expression level of PDX1 and secretion of insulin proteins were increased.
Results: Results showed that during time, the cell viability remained high at 87% at day 7. After 21 days, the differentiated beta-like cells in microcapsules were morphologically similar to primary beta cells. Evaluation of the expression of PDX1 and INS by quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) on days 7, 14 and 21 of differentiation exhibited the highest expression on day 14. At the protein level, the expression of these two pancreatic markers was observed after PDX1 transduction. Results showed that the intracellular and extracellular insulin levels in the cells receiving PDX1 is higher than the control group. The current data showed that encapsulation with alginate by PLL and PLO outer layer permitted to increase the microcapsules’ beta cell differentiation.
Conclusion: Encapsulate the transduced-MSCs with alginate can be applied in an in vivo model in order to do the further analysis.


1. Association AD. Diagnosis and classification of diabetes mellitus. Diabetes care. 2010; 33 Suppl 1: S62-S69.
2. Shaw JE, Sicree RA, Zimmet PZ. Global estimates of the prevalence of diabetes for 2010 and 2030. Diabetes Res Clin Pract. 2010; 87(1): 4-14.
3. Kuncorojakti S, Rodprasert W, Yodmuang S, Osathanon T, Pavasant P, Srisuwatanasagul S, et al. Alginate/Pluronic F127-based encapsulation supports viability and functionality of human dental pulp stem cell-derived insulin-producing cells. J Biol Eng. 2020; 14(1): 1-15.
4. Pokrywczynska M, Krzyzanowska S, Jundzill A, Adamowicz J, Drewa T. Differentiation of stem cells into insulin-producing cells: current status and challenges. Arch Immunol Ther Exp (Warsz). 2013; 61(2): 149-158.
5. Yu S, Li C, Hou Xg, Hou Wk, Dong JJ, Lei S, et al. Differentiation of bone marrow-derived mesenchymal stem cells from diabetic patients into insulin-producing cellsin vitro. Chin Med J. 2007; 120(9): 771-776.
6. Yakhkeshi S, Rahimi S, Sharafi M, Hassani SN, Taleahmad S, Shahverdi A, et al. In vitro improvement of quail primordial germ cell expansion through activation of TGF-beta signaling pathway. J Cell Biochem. 2018; 119(6): 4309-4319.
7. Schiesser JV, Wells JM. Generation of β cells from human pluripotent stem cells: are we there yet? Ann N Y Acad Sci. 2014; 1311: 124-137.
8. Chmielowiec J, Borowiak M. In vitro differentiation and expansion of human pluripotent stem cell-derived pancreatic progenitors. Rev Diabet Stud. 2014; 11(1): 19-34.
9. Moshtagh PR, Emami SH, Sharifi AM. Differentiation of human adipose-derived mesenchymal stem cell into insulin-producing cells: an in vitro study. J Physiol Biochem. 2013; 69(3): 451-458.
10. Hu J, Yu X, Wang Z, Wang F, Wang L, Gao H, et al. Long term effects of the implantation of Wharton’s jelly-derived mesenchymal stem cells from the umbilical cord for newly-onset type 1 diabetes mellitus. Endocr J. 2013; 60(3): 347-357.
11. Jiang R, Han Z, Zhuo G, Qu X, Li X, Wang X, et al. Transplantation of placenta-derived mesenchymal stem cells in type 2 diabetes: a pilot study. Front Med. 2011; 5(1): 94-100.
12. Andersen T, Auk-Emblem P, Dornish M. 3D cell culture in alginate hydrogels. Microarrays. 2015; 4(2): 133-161.
13. Kin K, Yasuhara T, Kameda M, Tomita Y, Umakoshi M, Kuwahara K, et al. Cell encapsulation enhances antidepressant effect of the mesenchymal stem cells and counteracts depressive-like behavior of treatment-resistant depressed rats. Mol Psychiatry. 2020; 25(6): 1202-1214.
14. Sarker B, Rompf J, Silva R, Lang N, Detsch R, Kaschta J, et al. Alginate-based hydrogels with improved adhesive properties for cell encapsulation. Int J Biol Macromol. 2015; 78: 72-78.
15. Sun J, Tan H. Alginate-based biomaterials for regenerative medicine applications. Materials. 2013; 6(4): 1285-1309.
16. Choi S, Kim JH, Ha J, Jeong BI, Jung YC, Lee GS, et al. Intraarticular injection of alginate-microencapsulated adipose tissuederived mesenchymal stem cells for the treatment of osteoarthritis in rabbits. Stem Cells Int. 2018; 2018.
17. Strand BL, Coron AE, Skjak-Braek G. Current and future perspectives on alginate encapsulated pancreatic islet. Stem Cells Transl Med. 2017; 6(4): 1053-1058.
18. Lim F, Sun AM. Microencapsulated islets as bioartificial endocrine pancreas. Science. 1980; 210(4472): 908-910.
19. Kumar SR, Markusic DM, Biswas M, High KA, Herzog RW. Clinical development of gene therapy: results and lessons from recent successes. Mol Ther Methods Clin Dev. 2016; 3: 16034.
20. Tsokos GC, Nepom GT. Gene therapy in the treatment of autoimmune diseases. J Clin Invest. 2000; 106(2): 181-183.
21. Allahverdi A, Abroun S, Jafarian A, Soleimani M, Taghikhani M, Eskandari F. Differentiation of human mesenchymal stem cells into insulin producing cells by using a lentiviral vector carrying PDX1. Cell J. 2015; 17(2): 231.
22. Gerace D, Martiniello-Wilks R, Simpson A. Diabetes reversal via gene transfer: building on successes in animal models. Res Rep Endocr Disord. 2015; 5: 15-29
23. Varaa N, Azandeh S, Khodabandeh Z, Gharravi AM. Wharton’s jelly mesenchymal stem cell: Various protocols for isolation and differentiation of hepatocyte-like cells; narrative review. Iran J Med Sci. 2019; 44(6): 437-448.
24. Noguchi H, Xu G, Matsumoto S, Kaneto H, Kobayashi N, Bonner-Weir S, et al. Induction of pancreatic stem/progenitor cells into insulin-producing cells by adenoviral-mediated gene transfer technology. Cell Transplant. 2006; 15(10): 929-938.
25. Kanafi MM, Ramesh A, Gupta PK, Bhonde RR. Dental pulp stem cells immobilized in alginate microspheres for applications in bone tissue engineering. Int Endod J. 2014; 47(7): 687-697.
26. Gabr MM, Zakaria MM, Refaie AF, Abdel-Rahman EA, Reda AM, Ali SS, et al. From human mesenchymal stem cells to insulinproducing cells: comparison between bone marrow-and adipose tissue-derived cells. Biomed Res Int. 2017; 2017.
27. Fiorina P, Voltarelli J, Zavazava N. Immunological applications of stem cells in type 1 diabetes. Endocr Rev. 2011; 32(6): 725-754.
28. El-Badawy A, El-Badri N. Clinical efficacy of stem cell therapy for diabetes mellitus: a meta-analysis. PLoS One. 2016; 11(4): e0151938.
29. Czubak P, Bojarska-Junak A, Tabarkiewicz J, Putowski L. A modified method of insulin producing cells’ generation from bone marrow-derived mesenchymal stem cells. J Diabetes Res. 2014; 2014: 628591.
30. Ranjbaran H, Abediankenari S, Amiri MM. Enhanced differentiation of whartonâ s jelly-derived mesenchymal stem cells in insulin-producing cells by the extract of nigella sativa seeds. Iran Red Crescent Med J. 2018; 20(3).
31. Sharma A, Rani R. Do we really need to differentiate mesenchymal stem cells into insulin-producing cells for attenuation of the autoimmune responses in type 1 diabetes: immunoprophylactic effects of precursors to insulin-producing cells. Stem Cell Res Ther. 2017; 8(1): 167.
32. Rahmati S, Alijani N, Kadivar M. In vitro generation of glucose-responsive insulin producing cells using lentiviral based pdx-1 gene transduction of mouse (C57BL/6) mesenchymal stem cells. Biochem Biophys Res Commun. 2013; 437(3): 413-419.
33. Soltanian A, Ghezelayagh Z, Mazidi Z, Halvaei M, Mardpour S, Ashtiani MK, et al. Generation of functional human pancreatic organoids by transplants of embryonic stem cell derivatives in a 3Dprinted tissue trapper. J Cell Physiol. 2019; 234(6): 9564-9576.
34. Lima MJ, Muir KR, Docherty HM, McGowan NW, Forbes S, Heremans Y, et al. Generation of functional beta-like cells from human exocrine pancreas. PLoS One. 2016; 11(5): e0156204.
35. Barati G, Nadri S, Hajian R, Rahmani A, Mostafavi H, Mortazavi Y, et al. Differentiation of microfluidic-encapsulated trabecular meshwork mesenchymal stem cells into insulin producing cells and their impact on diabetic rats. J Cell Physiol. 2019; 234(5): 6801-6809.
36. Alagpulinsa DA, Cao JJ, Driscoll RK, Sîrbulescu RF, Penson MF, Sremac M, et al. Alginate-microencapsulation of human stem cell–derived β cells with CXCL 12 prolongs their survival and function in immunocompetent mice without systemic immunosuppression. Am J Transplant. 2019; 19(7): 1930-1940.
37. Khatab S, Leijs MJ, van Buul G, Haeck J, Kops N, Nieboer M, et al. MSC encapsulation in alginate microcapsules prolongs survival after intra-articular injection, a longitudinal in vivo cell and bead integrity tracking study. Cell Biol Toxicol. 2020; 36(6): 553-570.
38. Duruksu G, Polat S, Kayiş L, Gürcan NE, Gacar G, Yazir Y. Improvement of the insulin secretion from beta cells encapsulated in alginate/poly-L-histidine/alginate microbeads by platelet-rich plasma. Turk J Biol. 2018; 42(4): 297-306.
39. Tsai PJ, Wang HS, Lin GJ, Chou SC, Chu TH, Chuan WT, et al. Undifferentiated Wharton’s jelly mesenchymal stem cell transplantation induces insulin-producing cell differentiation and suppression of T-cell-mediated autoimmunity in nonobese diabetic mice. Cell Transplant. 2015; 24(8): 1555-1570.
40. De Mesmaeker I, Robert T, Suenens KG, Stangé GM, Van Hulle F, Ling Z, et al. Increase functional β-cell mass in subcutaneous alginate capsules with porcine prenatal islet cells but loss with human adult islet cells. Diabetes. 2018; 67(12): 2640-2649.