Culture of endocrine pancreatic cells in protein-free, chemically defined media
Autor: | Fabienne Kinard, L. De Clercq, J. J. Hoet, B. Amory, Claude Remacle, B. Billen |
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Rok vydání: | 1990 |
Předmět: |
medicine.medical_specialty
medicine.medical_treatment Clinical Biochemistry Plant Science Biology Islets of Langerhans Leucine Internal medicine Insulin Secretion medicine Animals Insulin Fibroblast Cells Cultured chemistry.chemical_classification geography geography.geographical_feature_category Rats Inbred Strains Cell Biology equipment and supplies Islet Culture Media Rats Chemically defined medium Microscopy Electron medicine.anatomical_structure Endocrinology Glucose chemistry Cell culture Transferrin Collagenase Secretory Rate Fetal bovine serum Cell Division Biotechnology Developmental Biology medicine.drug |
Zdroj: | In vitro cellulardevelopmental biology : journal of the Tissue Culture Association. 26(10) |
ISSN: | 0883-8364 |
Popis: | Cell suspensions prepared by collagenase digestion of pancreata obtained from 21.5-d-old rat fetuses were preincubated in RPMI medium containing 10% fetal bovine serum (FBS), to ensure cell adhesion. Twenty hours later, this medium was replaced by a chemically defined medium. Dulbecco's modified Eagle's (DME)-F12 was used alone or supplemented with various combinations of transferrin, sodium selenite, or Ultroser G. The evolution of the culture and the islet ultrastructure were similar in defined and serum-containing media. However, in the defined medium, the neoformed islets seemed less numerous, and the fibroblast layer less dense, when compared to the RPMI + 10% FBS control medium. At Day 7, in defined media, the total insulin content per dish was half that of control cultures. None of the tested additives improved the yield of the cultures. The fractional insulin release per day was elevated in defined media. In subsequent incubations, glucose and leucine stimulated insulin release in a way characteristic of these cells of fetal origin. The labeling index of islet cells cultured in DME-F12 reached 10.7%, which is not far from that observed in RPMI + 10% FBS. Such a defined medium is useful to study B cell physiology, avoiding the possible interaction of serum components with substances to be tested. |
Databáze: | OpenAIRE |
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