title: Pancreatic tissue resident mesenchymal stromal cell (MSC)-like cells as a source of in vitro islet neogenesis.
abstract: Insufficient β-cell mass is a common denominator for both type 1 and type 2 diabetes. In vitro generation of β-cells from islet precursor cells, exocrine cells or ductal epithelia provide an alternative source of insulin-producing cells. However the presence of multipotent precursor cells within the pancreas is also deliberated. In this study we isolated mesenchymal stromal cell (MSC)-like cells from adult mouse pancreas by collagenase digestion. We used Knockout DMEM for our isolation procedure and the floating islets and acini were removed after 48 h. This strategy permitted the adhesion of stromal cells with typical mesenchymal morphology. These cells not only expressed MSC-specific markers like Sca-1, CD90.2, CD73, and CD44 but also generated osteocytes, adipocytes, and neurons when induced with specific growth media. Upon exposure to islet differentiation serum-free cocktail a significant upregulation of pancreatic markers like Nkx2.2, Nkx6.1, Pdx1, insulin, and somatostatin was seen. The differentiated islet-like cell aggregates (ICAs) secreted insulin which increased over the days in culture in presence of basal glucose levels. Taken together, our data strongly indicate that there is a tissue-resident precursor population within the pancreas that can be exploited for islet neogenesis in vitro.
authors.affiliation: Manipal Institute of Regenerative Medicine (MIRM), Manipal University, Bangalore, India.
authors.forename: Renjitha
authors.initials: R
authors.lastname: Gopurappilly
authors.forename: Vijay
authors.initials: V
authors.lastname: Bhat
authors.forename: Ramesh
authors.initials: R
authors.lastname: Bhonde
mesh_terms: D015720:5'-Nucleotidase / Q000378:metabolism; D000818:Animals; D000950:Antigens, Ly / Q000378:metabolism; D002453:Cell Cycle / Q000502:physiology; D002454:Cell Differentiation / Q000502:physiology; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D000090322:Homeobox Protein Nkx-2.2; D018398:Homeodomain Proteins / Q000378:metabolism; D018960:Hyaluronan Receptors / Q000378:metabolism; D007150:Immunohistochemistry; D007328:Insulin / Q000378:metabolism; D007515:Islets of Langerhans / Q000166:cytology* / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D010179:Pancreas / Q000166:cytology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013004:Somatostatin / Q000378:metabolism; D018800:Thy-1 Antigens / Q000378:metabolism; D015534:Trans-Activators / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism; D029961:Zebrafish Proteins
chemical_list: D000950:Antigens, Ly; D000090322:Homeobox Protein Nkx-2.2; D018398:Homeodomain Proteins; D018960:Hyaluronan Receptors; D007328:Insulin; C074650:Ly6a protein, mouse; D008565:Membrane Proteins; C000715627:Nkx2-2 protein, mouse; C491328:Nkx6-1 protein, mouse; D018800:Thy-1 Antigens; D015534:Trans-Activators; D014157:Transcription Factors; D029961:Zebrafish Proteins; C000715630:nkx2.2b protein, zebrafish; C084010:pancreatic and duodenal homeobox 1 protein; D013004:Somatostatin; D015720:5'-Nucleotidase
keywords: INSULIN SECRETION; ISLET NEOGENESIS; MESENCHYMAL STROMAL CELL DIFFERENTIATION; β-CELL ENRICHMENT