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Ultrafiltration and Injection of Islet Regenerative Stimuli Secreted by Pancreatic Mesenchymal Stromal Cells
- Source :
- Stem Cells and Development. 30:247-264
- Publication Year :
- 2021
- Publisher :
- Mary Ann Liebert Inc, 2021.
-
Abstract
- The secretome of mesenchymal stromal cells (MSCs) is enriched for biotherapeutic effectors contained within and independent of extracellular vesicles (EVs) that may support tissue regeneration as an injectable agent. We have demonstrated that the intrapancreatic injection of concentrated conditioned media (CM) produced by bone marrow MSC supports islet regeneration and restored glycemic control in hyperglycemic mice, ultimately providing a platform to elucidate components of the MSC secretome. Herein, we extend these findings using human pancreas-derived MSC (Panc-MSC) as "biofactories" to enrich for tissue regenerative stimuli housed within distinct compartments of the secretome. Specifically, we utilized 100 kDa ultrafiltration as a simple method to debulk protein mass and to enrich for EVs while concentrating the MSC secretome into an injectable volume for preclinical assessments in murine models of blood vessel and islet regeneration. EV enrichment (EV+) was validated using nanoscale flow cytometry and atomic force microscopy, in addition to the detection of classical EV markers CD9, CD81, and CD63 using label-free mass spectrometry. EV+ CM was predominately enriched with mediators of wound healing and epithelial-to-mesenchymal transition that supported functional regeneration in mesenchymal and nonmesenchymal tissues. For example, EV+ CM supported human microvascular endothelial cell tubule formation in vitro and enhanced the recovery of blood perfusion following intramuscular injection in nonobese diabetic/severe combined immunodeficiency mice with unilateral hind limb ischemia. Furthermore, EV+ CM increased islet number and β cell mass, elevated circulating insulin, and improved glycemic control following intrapancreatic injection in streptozotocin-treated mice. Collectively, this study provides foundational evidence that Panc-MSC, readily propagated from the subculture of human islets, may be utilized for regenerative medicine applications.
- Subjects :
- 0301 basic medicine
Ultrafiltration
Mice, SCID
Biology
Microscopy, Atomic Force
Proteomics
Extracellular vesicles
Regenerative medicine
Streptozocin
Biological Factors
Extracellular Vesicles
03 medical and health sciences
0302 clinical medicine
Mice, Inbred NOD
medicine
Animals
Humans
Regeneration
Pancreas
Cells, Cultured
Secretome
geography
geography.geographical_feature_category
Mesenchymal stem cell
Endothelial Cells
Mesenchymal Stem Cells
Cell Biology
Hematology
Islet
Cell biology
Transplantation
Ultrafiltration (renal)
030104 developmental biology
medicine.anatomical_structure
Culture Media, Conditioned
Hyperglycemia
Blood Vessels
030217 neurology & neurosurgery
Developmental Biology
Subjects
Details
- ISSN :
- 15578534 and 15473287
- Volume :
- 30
- Database :
- OpenAIRE
- Journal :
- Stem Cells and Development
- Accession number :
- edsair.doi.dedup.....e9f0e2113e55edca03b036f6823149a3