Decreasing Loss of Cryopreserved-Thawed Rat Islets by Coculture with Sertoli Cells

Abstract Objective Cryopreserved-thawed rat islets were cocultured with Sertoli cells to examine whether they could decrease the loss and improve islet function. Methods Islets and Sertoli cells were harvested from the pancreas and the testis of Sprague-Dawley rats, respectively. Cryopreserved, stor...

Full description

Saved in:
Bibliographic Details
Published in:Transplantation proceedings 2012-06, Vol.44 (5), p.1423-1428
Main Authors: Li, Y, Xue, W.-J, Wang, X.-H, Tian, X.-H, Liu, H.-B, Feng, X.-S, Ding, X.-M, Tian, P.-X, Pan, X.-M, Ding, C.-G, Zheng, J
Format: Article
Language:English
Subjects:
Animals
Apoptosis
Apoptosis Regulatory Proteins - genetics
Apoptosis Regulatory Proteins - metabolism
Biological and medical sciences
Blotting, Western
Cell Shape
Cells, Cultured
Coculture Techniques
Cryopreservation
Diabetes Mellitus, Experimental - blood
Diabetes Mellitus, Experimental - surgery
Enzyme-Linked Immunosorbent Assay
Fundamental and applied biological sciences. Psychology
Fundamental immunology
Glucose - metabolism
Graft Survival
Insulin - metabolism
Intercellular Signaling Peptides and Proteins - metabolism
Islets of Langerhans - metabolism
Islets of Langerhans - pathology
Islets of Langerhans Transplantation
Male
Medical sciences
Mice
Mice, Nude
Paracrine Communication
Rats
Rats, Sprague-Dawley
Reverse Transcriptase Polymerase Chain Reaction
Sertoli Cells - metabolism
Surgery
Surgery (general aspects). Transplantations, organ and tissue grafts. Graft diseases
Time Factors
Tissue Culture Techniques
Tissue Survival
Tissue, organ and graft immunology
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Abstract Objective Cryopreserved-thawed rat islets were cocultured with Sertoli cells to examine whether they could decrease the loss and improve islet function. Methods Islets and Sertoli cells were harvested from the pancreas and the testis of Sprague-Dawley rats, respectively. Cryopreserved, stored islets were thawed and divided into groups of coculture with Sertoli cells versus single cells. We measured islets recovery rate and function. Apoptotic-related proteins and gene expressions were detected by Western blot and reverse-transcriptase polymerase chain reaction. Soluble factors secreted by Sertoli cells in to the supernate were detected by enzyme-linked immunosorbent assay. We compared islet graft survival times in diabetic mice. Results In contrast to the single culture controls, thawed islets cocultured with Sertoli cells exhibited improved morphology. Recovery rates and insulin secretion were significantly higher among coculture cells. Four soluble factors were detected in supernates from Sertoli cell cultures including transforming growth factor-β, insulin-like growth factor-1, epidermal growth factor, and basic fibroblast growth factor. Expression of proapoptotic Bax and caspase 3, 7 were down-regulated while that of antiapoptotic Bcl-2 was up-regulated. Cotransplantation with Sertoli cells significantly prolonged islet graft survival. Conclusion These results suggested that coculture with Sertoli cells significantly improved islet yields and function after thawing and depressed islet apoptosis.
ISSN:0041-1345
1873-2623
DOI:10.1016/j.transproceed.2012.01.143