Production of insulin producing cells from cord blood mesenchymal stem cells and their potential in cell therapy
Introduction: Mesenchymal stem cells (MSCs) were described as adherent cells with a fibroblast-like appearance, have a great capacity for self-renewal while maintaining their multipotency and differentiation into multiple tissues in vivo and in vitro. Methods: MSCs were isolated from cord blood of Sudanese donors using Ficoll-Hypaque gradient density protocol, and differentiate into β- like cells using 3-step protocol. STZ induced diabetic rats were injected intraperitoneally with the differentiated islet β- like cells and blood glucose levels were monitored for seven days. Results: The adherent cell appeared round and sphere after one-week of incubation, and the fibroblast-like colony was strongly attached after three weeks of seeding. The phenotyping of cells showed positivity for CD13, and negativity for CD34, CD45 and HLADR. MSCs were induced into islet-like cells using a 3-step (15-days) protocol. The differentiated cells showed positive diathizone stain and positive imuno-reactivity to anti-human insulin antibody. Secretion of insulin by insulin-producing cells showed positive result with >3.4 u/ml scale reading in high glucose concentration medium. After one-week of transplantation the level of blood glucose was reduced from 410 to 225 mg/dl in the experimental rat. Conclusion: Human UCB-MSCs can be differentiated into insulin-secreting cells invitro, and are able to produce and secrete insulin in response to high glucose concentration in vivo and in vitro.
Keywords: Cord blood, Mesenchymal stem cell, islets β-like cells
2- Hoogduijn, M. Are mesenchymal stromal cells immune cells. Arthritis Research and Therapy 2015; 17: 88.
3- Brighton, C.T, Hunt, R.M. Early histological and ultra-structural changes in medullary fracture callus. The Journal of Bone and Joint Surgery 1991; 73: 832-847.
4- Hematti, P. Mesenchymal stromal cells and fibroblasts: a case of mistaken identity. Cytotherapy 2012; 14: 516-521.
5- Zhao, Q.J., A., Ren, H.Y., Han, Z.C. Mesenchymal stem cells: Immunomodulatory capability and clinical potential in immune diseases. Journal of Cellular Immunotherapy2016; 2:3-20.
6- Pittenger, M.F., Mackay, A.M., Beck, S.C., Jaiswal, R.K., Douglas, R., Mosca, J.D., Mooman, M.A., Simonetti, D.W., Craig, S., Marshak, D.R. Multilineage potential of adult human mesenchymal stem cells. Science1999; 284:143-147.
7- Flynn, A., Barry, F., O'Brien, T. UC blood-derived mesenchymal stromal cells an overview. Cytotherapy 2007;9: 717-726
8- Hadar, A.Z., Lazarovici, P., Nagler, A. Tissue regeneration potential in human umbilical cord blood. Best Practice and Research Clinical Hematology2010; 23:291-303.
9- Malgieri, A., Kantzari, E., Patrizi, M.P., Gambardella, S. Bone marrow and umbilical cord blood human mesenchymal stem cells: state of the art. International Journal of Clinical and Experimental Medicine 2010; 3:248-269.
10- Sabatini, F., Petecchia, L., Tavian, M., de Villeroche, V. J., Rossi, G. A. Brouty-Boye, D. Human bronchial fibroblasts exhibit a mesenchymal stem cell phenotype and multilineage differentiating potentialities. Laboratory Investigation2005; 85:962-971
11- Gao, F., Wu, D.Q., Hu, Y.H., Jin, G.X., Li, G.D., Sun, T.W., Li, F.J. In vitro cultivation of islet-like cell clusters from human umbilical cord blood-derived mesenchymal stem cells. Translational Research 2008; 151:293-302. https://doi.org/10.1016/j.trsl.2008.03.003
12- Bieback, K., kinzebach, S., karagianni, M. Translating research into clinical scale manufacturing of mesenchymal stromal cells. Stem cells International 2011. Article ID 193519, 11 pages doi:10.4061/2010/193519
13- Zeddou,M.,Briquette,A.,Relic,B.,Jossr,C.,Malaise,M.G.,Gothot,A.,Lachanteur,C.,Beguin,Y. The umbilical cord matrix is a better source of mesenchymal stem cells (MSC) than the umbilical cord blood. Cell Biology International 2010; 34:693-701.
14- Wankhade, U.D., Shen, M., Kolhe, R., Fulzele, S. Advances in Adipose-Derived Stem Cells Isolation, Characterization, and Application in Regenerative Tissue Engineering. Stem Cells International 2016. ID 3206807, 9
15- Kern, S., Eichler, H., Stove, J., Klater, H., Bieback, K. Comparative analysis of mesenchymal stem cells from bone marrow, umbilical cord blood, or adipose tissue. Stem cells2006; 20:294-301.
16- Tondreau, T., Meuleman, N., Delforge, A., Dejeneffe, M., Leroy, R., Massy, M., Mortier, C., Bron, D., Lagneaux, L. Mesenchymal Stem Cells Derived from CD133-Positive Cells in Mobilized Peripheral Blood and Cord Blood Proliferation, Oct4 Expression, and Plasticity. Stem Cells 2005; 23:1105-1112.
17- Zhang, X.Y., Block, N.L., Schally, A.V. Cell based therapy for treatment of diabetes mellitus: can the agonists of growth hormone releasing hormone make a contribution. International Journal of Stem Cell Research and Therapy 2016; 2(1):469-570.
18- Yang, L., Li, S., Hatch, H., Ahrens, K., Comelus, J.G., Petersen, B.E., Peck, A.B. In vitro trans-differentiation of adult hepatic stem cells into pancreatic endocrine hormone-producing cells. Proceeding of the National Academy of Sciences of the United State of America 2002; 99: 8078-8083.
19- Cras-Meneur, C., Elghazi, L., Czernichow, P., Scharfmann, R. Epidermal growth factor increases undifferentiated pancreatic embryonic cells in vitro a balance between proliferation and differentiation. Diabetes 2001; 50:1571-1579.
20- . Gao, R., Ustinov, J., Pulkkinen, M.A., Lundin, K., Korsgren, O., Otonkoski, T. Characterization of endocrine progenitor cells and critical factors for their differentiation in human adult pancreatic cell culture. Diabetes 2003; 52:15
21- Chen, L.B., Jiang, X.B., Yang, L. Differentiation of rat marrow mesenchymal stem cells into pancreatic islet beta-cells. World Journal of Gastroenterology2004; 10:3016-3020.
22- Parekh, V.S., Joglekar, M.V., Hardikar, A.A. Differentiation of human umbilical cord blood-derived mononuclear cells to endocrine pancreatic lineage. Differentiation2209; 78:232- 240.
23- Tsai P.J., Wang, H.S., Shyr, Y.M., Weng, Z.C., Tai, L.C., Shyu, J.F., Chen, T.H. Transplantation of insulin-producing cells from umbilical cord mesenchymal stem cells for the treatment of streptozotocin induced diabetic rats.Journal of Biomedical Science 2012; 19-47. doi:10.1186/1423-0127-19-47.
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
Authors who publish with this journal agree to the following terms:
- Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported License. that allows others to share the work with an acknowledgment of the work's authorship and initial publication in this journal.
- Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgment of its initial publication in this journal.
- Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (SeeÂ The Effect of Open Access).