Tailored generation of insulin producing cells from canine mesenchymal stem cells derived from bone marrow and adipose tissue

Abstract

The trend of regenerative therapy for diabetes in human and veterinary practices has conceptually been proven according to the Edmonton protocol and animal models. Establishing an alternative insulin-producing cell (IPC) resource for further clinical application is a challenging task. This study investigated IPC generation from two practical canine mesenchymal stem cells (cMSCs), canine bone marrow-derived MSCs (cBM-MSCs) and canine adipose-derived MSCs (cAD-MSCs). The results illustrated that cBM-MSCs and cAD-MSCs contain distinct pancreatic differentiation potential and require the tailor-made induction protocols. The effective generation of cBM-MSC-derived IPCs needs the integration of genetic and microenvironment manipulation using a hanging-drop culture of PDX1-transfected cBM-MSCs under a three-step pancreatic induction protocol. However, this protocol is resource- and time-consuming. Another study on cAD-MSC-derived IPC generation found that IPC colonies could be obtained by a low attachment culture under the three-step induction protocol. Further, Notch signaling inhibition during pancreatic endoderm/progenitor induction yielded IPC colonies through the trend of glucose-responsive C-peptide secretion. Thus, this study showed that IPCs could be obtained from cBM-MSCs and cAD-MSCs through different induction techniques. Also, further signaling manipulation studies should be conducted to maximize the protocol’s efficiency.

In vitro generation of transplantable insulin-producing cells from canine adipose-derived mesenchymal stem cells

“…Transcription of Hes1 gene inhibits the promotion of Ngn3 gene 99 , while Ngn3 expression is essentially required for endocrine cell development 98 . Our previous studies proved that the inhibition of Notch signaling during PE induction benefits IPC generation from MSCs 38 , 69 . Although RA could also suppress the Hes1 expression indirectly 93 , further analyses are still required.…”

“…In this study, cAD-MSC induction protocol toward mature IPCs and the enriched medium for preserving AGLPA-encapsulated cAD-MSC-derived IPCs were established to produce RTU IPCs. The cAD-MSCs were isolated, cultured, and expanded following our previous report 38 . These isolated cAD-MSCs were then defined following the International Society for Cellular Therapy (ISCT) 39 , they presented fibroblast-like cell sharp, adherent to culture plastic.…”

“…Furthermore, MSCs are widely used in clinical trials 68 , thus MSCs are offered as a good candidate for IPC generation 16 . The MSC-based protocols mainly comprise of three differentiation stages 16 , 38 , 69 – 71 . The different stages for MSCs are DE, PE progenitors, and IPC maturation.…”

“…The different stages for MSCs are DE, PE progenitors, and IPC maturation. Although IPCs could be produced from cAD-MSCs with the simple three-stepwise protocol 69 , 70 , their function is still hamperred 38 . Here, we illustrated that the modified three-stepwise protocol could improve the in vitro functional IPC differentiation from cAD-MSCs.…”

“…As the first step of IPC-derived MSCs, DE formation is a prerequisite for generating efficient pancreatic lineage from MSCs 72 . In previous studies, Activin A and Chir99021 were known as the potential small molecules on DE development 38 , 69 , 70 , 73 . Although the independent utilization of Chir99021 and Activin A could induce DE from stem cells 74 – 77 , the cooperation of Chir99021 and Activin A could optimize for inducing DE cells 76 , 77 .…”