Therefore, this study focused on establishing a protocol for cAD-MSCs induction toward mature IPCs in vitro.
In types of cMSCs, canine adipose-derived MSCs (cAD-MSCs) are an accessible candidate and possess the potency for IPC differentiation 22, 23. To fabricate the effective cMSC-derived IPCs, it is essential to advance the current differentiation protocols.
Although a minority of research on IPCs has originated from canine MSCs (cMSCs) 21, 22, 23, these cMSC-derived IPCs are still functionally inadequate and morphologically circumscribed. The capacity of human MSCs (hMSCs) differentiated into IPCs as well as their clinical accomplishment has been shown in many previous studies 17, 18, 19, 20. Meanwhile, MSCs possess immune-privileged and highly plastic abilities, this allows MSCs to be a wonderful and safe choice for IPC generation 16. Nevertheless, ESCs have encountered ethical issues, while reprogramming of iPSCs can cause teratoma formation and side effects of pluripotency-induced viral transgenes might be unsafe for clinical applications 16. To solve the restrictions of the Edmonton protocol, the tendency of regenerative medicine production in which insulin-producing cells (IPCs) derived from stem cells has been a promising candidate 12, 15.Ĭurrent in vitro IPCs are generated by embryonic stem cells (ESCs), induced pluripotent stem cells (iPSCs) and mesenchymal stem cells (MSCs). Although pancreatic islet transplantation can surmount the impediments of insulin therapy, the lack of donor islet source and the immune reactivity of recipients exist as two main obstacles of this method 11, 12, 14.
#STEM CELL TREATMENT FOR DIABETES TYPE 1 COST TRIAL#
In 2000, a trial of islet transplantation was performed successfully according to “Edmonton protocol”, thus this method introduced as an alternative approach for treating long-term hyperglycemia with insulin independence 11, 12, 13. Insulin therapy has been clinically well-established to manage T1DM in both dogs and humans, however, adverse events and disadvantages have also been periodically reported 8, 9, 10. By pathophysiological diagnosis, canine DM has been mostly concerned with beta cell deficiency by latent autoimmunity, which is considered as similar with human type 1 DM (T1DM) 6, 7. An epidemiological study in the United States reported a 32 percent increase in canine diabetes between 20, and the data kept on rising by 47.7 percent from 2011 to 2016 5. DM is one of the common endocrine diseases diagnosed in the canine family besides human beings 4. Together, the encapsulation of cAD-MSC-derived IPCs and the cultivation with VSCBIC-1 enrich the maturation of generated IPCs.ĭiabetes mellitus (DM) is a complex metabolic disorder characterized by a chronic presence of hyperglycemia and glycosuria as the results of insulin deficiency or impaired insulin response to target tissues 1, 2, 3. Conclusion, the modulated three-stepwise protocol generates the functional IPCs. Afterward, the maintenance of ALGPA-encapsulated cAD-MSC-derived IPCs with VSCBIC-1, a specialized medium, enhanced IPC properties. The last step of IPC production, the combination of taurine, nicotinamide, Glp-1, forskolin, PI3K inhibitor, and TGFβ inhibitor, yielded efficiently functional IPCs from PE precursors. The second step for pancreatic endocrine (PE) progenitor induction from DE indicated that the treatment with taurine, retinoic acid, FGF2, EGF, TGFβ inhibitor, dorsomorphin, nicotinamide, and DAPT showed the significant upregulation of the pancreatic endocrine precursor markers Pdx1 and Ngn3. The first step of definitive endoderm (DE) induction showed that the cooperation of Chir99021 and Activin A created the effective production of Sox17-expressed DE cells. IPCs were induced from cAD-MSCs with the modulated three-stepwise protocol. Subsequently, in vitro preservation of pluronic F127-coated alginate (ALGPA)-encapsulated cAD-MSC-derived IPCs was performed to verify ready-to-use IPCs. In this study, we established a useful protocol for generating IPCs from canine adipose mesenchymal stem cells (cAD-MSCs). Canine mesenchymal stem cells (cMSCs) have potential applications for regenerative therapy, including the generation of insulin-producing cells (IPCs) for studying and treating diabetes.
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