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Production, Characterization, and Function of Pseudoislets from Perinatal Canine Pancreas
We evaluated the cell composition and function of canine pancreatic pseudoislets (PIs) produced from 42- to 55-day-old fetuses, 1- to 21-day-old pups, and an adult dog pancreas. After mild collagenase treatment, partially digested tissues were cultured for 2–3 weeks. PI production started on culture...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
SAGE Publications
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6923560/ https://www.ncbi.nlm.nih.gov/pubmed/31450972 http://dx.doi.org/10.1177/0963689719869004 |
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author | Czernichow, P. Reynaud, K. Kerr-Conte, J. Furthner, E. Ravassard, P. |
author_facet | Czernichow, P. Reynaud, K. Kerr-Conte, J. Furthner, E. Ravassard, P. |
author_sort | Czernichow, P. |
collection | PubMed |
description | We evaluated the cell composition and function of canine pancreatic pseudoislets (PIs) produced from 42- to 55-day-old fetuses, 1- to 21-day-old pups, and an adult dog pancreas. After mild collagenase treatment, partially digested tissues were cultured for 2–3 weeks. PI production started on culture day 3, was marked for 6 to 9 days, and then stopped. PI production was greatest with the neonatal specimens, reaching about 12 million aggregates per litter (55-day-old fetus) or per pancreas (1-day-old pup). Cell composition at all stages was similar to that in adult pancreatic islets, with predominant β cells, scant α cells and, most importantly, presence of δ cells. Among pancreatic markers assessed by quantitative real-time PCR (qRT-PCR) mRNA assay, insulin showed the highest expression levels in PIs from newborn and adult pancreas, although these were more than 1000 times lower than in adult islets. Pdx1 mRNA expression was high in PIs from 55-day-old pancreases and was lower at later stages. Consistent with the qRT-PCR results, the insulin content was far lower than reported in adult dog pancreatic islets. However, insulin release by PIs from 1-day-old pups was demonstrated and was stimulated by a high-glucose medium. PIs were transplanted into euglycemic and diabetic SCID mice. In euglycemic animals, the transplant cell composition underwent maturation and transplants were still viable after 6 months. In diabetic mice, the PI transplants produced insulin and partially controlled the hyperglycemia. These data indicate that PIs can be produced ex vivo from canine fetal or postnatal pancreases. Although functional PIs can be obtained, the production yield is most likely insufficient to meet the requirements for diabetic dog transplantation without further innovation in cell culture amplification. |
format | Online Article Text |
id | pubmed-6923560 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | SAGE Publications |
record_format | MEDLINE/PubMed |
spelling | pubmed-69235602020-01-03 Production, Characterization, and Function of Pseudoislets from Perinatal Canine Pancreas Czernichow, P. Reynaud, K. Kerr-Conte, J. Furthner, E. Ravassard, P. Cell Transplant Original Articles We evaluated the cell composition and function of canine pancreatic pseudoislets (PIs) produced from 42- to 55-day-old fetuses, 1- to 21-day-old pups, and an adult dog pancreas. After mild collagenase treatment, partially digested tissues were cultured for 2–3 weeks. PI production started on culture day 3, was marked for 6 to 9 days, and then stopped. PI production was greatest with the neonatal specimens, reaching about 12 million aggregates per litter (55-day-old fetus) or per pancreas (1-day-old pup). Cell composition at all stages was similar to that in adult pancreatic islets, with predominant β cells, scant α cells and, most importantly, presence of δ cells. Among pancreatic markers assessed by quantitative real-time PCR (qRT-PCR) mRNA assay, insulin showed the highest expression levels in PIs from newborn and adult pancreas, although these were more than 1000 times lower than in adult islets. Pdx1 mRNA expression was high in PIs from 55-day-old pancreases and was lower at later stages. Consistent with the qRT-PCR results, the insulin content was far lower than reported in adult dog pancreatic islets. However, insulin release by PIs from 1-day-old pups was demonstrated and was stimulated by a high-glucose medium. PIs were transplanted into euglycemic and diabetic SCID mice. In euglycemic animals, the transplant cell composition underwent maturation and transplants were still viable after 6 months. In diabetic mice, the PI transplants produced insulin and partially controlled the hyperglycemia. These data indicate that PIs can be produced ex vivo from canine fetal or postnatal pancreases. Although functional PIs can be obtained, the production yield is most likely insufficient to meet the requirements for diabetic dog transplantation without further innovation in cell culture amplification. SAGE Publications 2019-08-26 2019-12 /pmc/articles/PMC6923560/ /pubmed/31450972 http://dx.doi.org/10.1177/0963689719869004 Text en © The Author(s) 2019 http://creativecommons.org/licenses/by/4.0/ This article is distributed under the terms of the Creative Commons Attribution 4.0 License (http://www.creativecommons.org/licenses/by/4.0/) which permits any use, reproduction and distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access pages (https://us.sagepub.com/en-us/nam/open-access-at-sage). |
spellingShingle | Original Articles Czernichow, P. Reynaud, K. Kerr-Conte, J. Furthner, E. Ravassard, P. Production, Characterization, and Function of Pseudoislets from Perinatal Canine Pancreas |
title | Production, Characterization, and Function of Pseudoislets from Perinatal
Canine Pancreas |
title_full | Production, Characterization, and Function of Pseudoislets from Perinatal
Canine Pancreas |
title_fullStr | Production, Characterization, and Function of Pseudoislets from Perinatal
Canine Pancreas |
title_full_unstemmed | Production, Characterization, and Function of Pseudoislets from Perinatal
Canine Pancreas |
title_short | Production, Characterization, and Function of Pseudoislets from Perinatal
Canine Pancreas |
title_sort | production, characterization, and function of pseudoislets from perinatal
canine pancreas |
topic | Original Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6923560/ https://www.ncbi.nlm.nih.gov/pubmed/31450972 http://dx.doi.org/10.1177/0963689719869004 |
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