Abstract
Umbilical cord blood (UCB)-derived mesenchymal stem cells (MSCs) are multipotent cells. They are able to differentiate into functional cells from not only mesoderm but also endoderm. Many researches showed that cells derived from fresh human UCB could transdifferentiate into insulin-secreting cells. In this study, transdifferentiating potential of cryopreserved human UCB-derived MSCs into insulin-secreting cell was investigated. Fresh human UCB was enriched the mononuclear cells by Ficoll–Paque centrifugation. The mononuclear cell population was cryopreserved in cryo-medium containing Iscove’s modified Dulbecco’s media (IMDM) with 10% DMSO at −196°C for 1 yr. After thawing, mononuclear cells were cultured to isolate MSCs in medium IMDM with 20% FBS supplemented with growth factors. At the fifth passages, MSCs were confirmed by flow cytometry about expression of CD13, CD14, CD34, CD45, CD166, and HLA-DR markers; after that, they were induced to differentiate into adipocytes and osteoblasts. After inducing with specific medium for islet differentiation, there were many clusters of cell like islet at day 14–28. Using real-time reverse transcription polymerase chain reaction (RT-PCR) to analyze the expression of functional genes, the result showed that Nestin, Pdx-1, Ngn3, Ils-1, Pax6, Pax4, Nkx2.2, Nkx6.1, Glut-2, Insulin genes expressed. The results showed that MSCs derived from banked cord blood can differentiate into functional pancreatic islet-like cells in vitro. If human MSCs, especially MSCs from banked cord blood of diabetes patients themselves can be isolated, proliferated, differentiated into functional pancreatic islet-like cells, and transplanted back into them (autologous transplantation), their high-proliferation potency and rejection avoidance will provide one promising therapy for diabetes.
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References
Assady S.; Maor G.; Amit M.; Itskovitz-Eldor J.; Skorecki K. L.; Tzukerman M. Insulin production by human embryonic stem cells. Diabetes 50: 1691–1697; 2001.
Bonner-Weir S.; Taneja M.; Weir G. C.; Tatarkiewicz K.; Song K. H.; Sharma A. et al. In vitro cultivation of human islets from expanded ductal tissue. Proc Natl Acad Sci USA 97: 7999–8004; 2000.
Davani S.; Marandin A.; Mersin N.; Royer B.; Kantelip B.; Herve P.; Etievent J. P.; Kantelip J. P. Mesenchymal progenitor cells differentiate into an endothelial phenotype, enhance vascular density, and improve heart function in a rat cellular cardiomyoplasty model. Circulation 108(Suppl 1): II253–258; 2003.
Deutsch G.; Jung J.; Zheng M.; Lora J.; Zaret K. S. A bipotential precursor population for pancreas and liver within the embryonic endoderm. Development 128: 871–881; 2001.
Gao F.; Wu D.; Hu Y.; Jin G. Extracellular matrix gel is necessary for in vitro cultivation of insulin producing cells from human umbilical cord blood derived mesenchymal stem cells. Chinese Medical Journal 121(9): 811–818; 2008.
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 52: 2007–2015; 2003.
Guz Y.; Nasir I.; Teitelman G. Regeneration of pancreatic beta cells from intra-islet precursor cells in an experimental model of diabetes. Endocrinology 142: 4956–4968; 2001.
Hardikar A. A.; Marcus-Samuels B.; Geras-Raaka E.; Raaka B. M.; Gershengorn M. C. Human pancreatic precursor cells secrete FGF2 to stimulate clustering into hormone-expressing islet-like cell aggregates. Proc Natl Acad Sci USA 100: 7117–7122; 2003.
Hori Y.; Rulifson I. C.; Tsai B. C.; Heit J. J.; Cahoy J. D.; Kim S. K. Growth inhibitors promote differentiation of insulin-producing tissue from embryonic stem cells. Proc Natl Acad Sci USA 99: 16105–16110; 2002.
Ianus A.; Holz G. G.; Theise N. D.; Hussain M. A. In vivo derivation of glucose-competent pancreatic endocrine cells from bone marrow without evidence of cell fusion. J Clin Invest 111: 843–850; 2003.
Jahr H.; Bretzel B. Insulin positive cells in vitro generated from rat bone marrow stromal cells. Transplant Proc 35: 2140–2141; 2003.
Jensen J.; Heller R. S.; Funder-Nielsen T.; Pedersen E. E.; Lindsell C.; Weinmaster G.; Madsen O. D.; Serup P. Independent development of pancreatic alpha- and beta-cells from neurogenin3-expressing precursors: a role for the notch pathway in repression of premature differentiation. Diabetes 49: 163–176; 2000.
Jiang Y.; Jahagirdar B. N.; Reinhardt R. L.; Schwartz R. E.; Keene C. D.; Ortiz-Gonzalez X. R.; Reyes M.; Lenvik T.; Lund T.; Blackstad M.; Du J.; Aldrich S.; Lisberg A.; Low W. C.; Largaespada D. A.; Verfaillie C. M. Pluripotency of mesenchymal stem cells derived from adult marrow. Nature 418: 41–49; 2002.
Kim D.; Gu Y.; Ishii M.; Fujimiya M.; Qi M.; Nakamura N.; Yoshikawa T.; Sumi S.; Inoue K. In vivo functioning and transplant table mature pancreatic islet-like cell clusters differentiated from embryonic stem cell. Pancreas 27: E34–E41; 2003.
Krause D. S.; Theise N. D.; Collector M. I.; Henegariu O.; Hwang S.; Gardner R.; Neutzel S.; Sharkis S. J. Multi-organ, multi-lineage engraftment by a single bone marrow-derived stem cells. Cell 105: 369–377; 2001.
Lee O. K.; Kuo T. K.; Chen W. M.; Lee K. D.; Hsieh S. L.; Chen T. H. Isolation of multipotent mesenchymal stem cells from umbilical cord blood. Blood 103: 1669–1675; 2004.
Leon-Quinto T.; Jones J.; Skoudy A.; Burcin M.; Soria B. In vitro directed differentiation of mouse embryonic stem cells into insulin producing cells. Diabetologia 47: 1442–1451; 2004.
Lumelsky N.; Blondel O.; Laeng P.; Velasco I.; Ravin R.; McKay R. Differentiation of embryonic stem cells to insulin secreting structures similar to pancreatic islets. Science 292: 1389–1394; 2001.
Petersen B. E.; Bowen W. C.; Patrene K. D.; Mars W. M.; Sullivan A. K.; Murase N.; Boggs S. S.; Greenberger J. S.; Goff J. P. Bone marrow as a potential source of hepatic oval cells. Science 284: 1168–1170; 1999.
Petropavlovskaia M.; Rosenberg L. Identification and characterization of small cells in the adult pancreas: potential progenitor cells? Cell Tissue Res 310: 51–58; 2002.
Pittenger M. F.; Mackay A. M.; Beck S. C.; Jaiswal R. K.; Douglas R.; Mosca J. D.; Moorman M. A.; Simonetti D. W.; Craig S.; Marshak D. R. Multilineage potential of adult human mesenchymal stem cells. Science 284: 143–147; 1999.
Reyes M.; Lund T.; Lenvik T.; Aguiar D.; Koodie L.; Verfaillie C. M. Purification and ex vivo expansion of postnatal human marrow mesodermal progenitor cells. Blood 98: 2615–2625; 2001.
Ryan E. A.; Lakey J. R. T.; Rajotte R. V.; Korbutt G. S.; Kin T.; Imes S. et al. Clinical outcomes and insulin secretion after islet transplantation with the Edmonton protocol. Diabetes 50: 710–719; 2001.
Schwartz R. E.; Reyes M.; Koodie L.; Jiang Y.; Blackstad M.; Lund T.; Lenvik T.; Johnson S.; Hu W. S.; Verfaillie C. M. Multipotent adult progenitor cells from bone marrow differentiate into functional hepatocyte-like cells. J Clin Invest 109: 1291–1302; 2002.
Shapiro A. M.; Lakey J. R.; Ryan E. A.; Korbutt G. S.; Toth E.; Warnock G. L. et al. Islet transplantation in seven patients with type 1 diabetes mellitus using a glucocorticoid-free immunosuppressive regime. N Engl J Med 343: 230–238; 2000.
Shiroi A.; Yoshikawa M.; Yokota H.; Fukui H.; Ishizaka S.; Tatsumi K.; Takahashi Y. Identification of insulin-producing cells derived from embryonic stem cells by zinc-chelating dithizone. Stem Cells 20: 284–292; 2002.
Soria B.; Roche E.; Berna G.; Leon-Quinto T.; Reig J. A.; Martin F. Insulin-secreting cells derived from embryonic stem cells normalize glycemia in streptozotocin-induced diabetic mice. Diabetes 49: 157–162; 2000.
Sun B.; Roh K.-H.; Lee S.-R.; Lee Y.-S.; Kang K.-S. Induction of human umbilical cord blood-derived stem cells with embryonic stem cell phenotypes into insulin producing islet-like structure. Biochemical and Biophysical Research Communications 354: 919–923; 2007.
Yang L.; Li S.; Hatch H.; Ahrens K.; Cornelius J. G.; Petersen B. E.; Peck A. B. In vitro trans-differentiation of adult hepatic stem cells into pancreatic endocrine hormon producing cells. Proc Natl Acad Sci USA 99: 8078–8083; 2002.
Zulewski H.; Abraham E. J.; Gerlach M. J.; Daniel P. B.; Moritz W.; Muller B. et al. Multipotential nestin-positive stem cells isolated from adult pancreatic islets differentiate ex vivo into pancreatic endocrine, exocrine, and hepatic phenotypes. Diabetes 50: 521–533; 2001.
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Editor: J. Denry Sato
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Phuc, P.V., Nhung, T.H., Loan, D.T.T. et al. Differentiating of banked human umbilical cord blood-derived mesenchymal stem cells into insulin-secreting cells. In Vitro Cell.Dev.Biol.-Animal 47, 54–63 (2011). https://doi.org/10.1007/s11626-010-9356-5
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DOI: https://doi.org/10.1007/s11626-010-9356-5