Direct Differentiation of Bone Marrow Mononucleated Cells Into Insulin-Producing Cells Using 4 Specific Soluble Factors

Abstract

Bone marrow-derived stem cells are self-renewing and multipotent adult stem cells that differentiate into several types of cells. Here, we investigated a unique combination of 4 differentiation-inducing factors (DIFs), including putrescine (Put), glucosamine (GlcN), nicotinamide, and BP-1-102, to develop a differentiation method for inducing mature insulin-producing cells (IPCs) and apply this method to bone marrow mononucleated cells (BMNCs) isolated from mice. BMNCs, primed with the 4 soluble DIFs, were differentiated into functional IPCs. BMNCs cultured under the defined conditions synergistically expressed multiple genes, including those for PDX1, NKX6.1, MAFA, NEUROG3, GLUT2, and insulin, related to pancreatic beta cell development and function. They produced insulin/C-peptide and PDX1, as assessed using immunofluorescence and flow cytometry. The induced cells secreted insulin in a glucose-responsive manner, similar to normal pancreatic beta cells. Grafting BMNC-derived IPCs under kidney capsules of mice with streptozotocin (STZ)-induced diabetes alleviated hyperglycemia by lowering blood glucose levels, enhancing glucose tolerance, and improving glucose-stimulated insulin secretion. Insulin- and PDX1-expressing cells were observed in the IPC-bearing graft sections of nephrectomized mice. Therefore, this study provides a simple protocol for BMNC differentiation, which can be a novel approach for cell-based therapy in diabetes mellitus.

Ameliorating and refining islet organoids to illuminate treatment and pathogenesis of diabetes mellitus

“…Endodermal ASCs, such as pancreatic ductal progenitor cells, PP cells, hepatic stem cells, and gastric stem cells, can directly differentiate into insulin-producing β-cells [ 14 , 27 , 28 ]. Additionally, cells from the mesodermal layer, such as bone marrow mesenchymal stem cells (BMSCs), adipose tissue stem cells (ADSCs), dental pulp stem cells, and ectodermal cells like amniotic epithelial cells, can also be induced to transdifferentiate into β-cells [ 29 , 30 ]. However, the efficiency of differentiating ASCs into β cells varies widely across studies, and their germline origin may be the main reason.…”

A Supportive Role of Mesenchymal Stem Cells on Insulin-Producing Langerhans Islets with a Specific Emphasis on The Secretome

“…Many protocols have been developed to differentiate MSCs into functional insulin-producing β-cells by exposing the cells to chemical and biological factors or by genetic manipulation introducing the PDX1 gene, which is a master regulator in pancreas organogenesis [ 232 , 437 , 440 , 441 , 442 , 443 , 444 , 445 , 446 , 447 , 448 , 449 , 450 , 451 , 452 , 453 , 454 , 455 , 456 , 457 , 458 , 459 ]. A common dominator for the different differentiation protocols is the sequential exposure of MSCs to different combinations of growth factors (e.g., EGF, bFGF, betacellulin, activin A, HGF, extendin-4, insulin) chemical compounds (e.g., nicotinamide), and B27 supplement (containing among others insulin, biotin, vitamin E, Vitamin A, selenium, putrescine, transferrin, catalase, superoxide dismutase, triodo-L-thyronine, linoleic and linolenic acids) for different time periods [ 441 , 442 ].…”