A serum-free defined culture system which maintains follicle-stimulating hormone responsiveness and differentiation of porcine granulosa cells.

Published

Journal Article

A serum-free defined culture system has been developed that maintains follicle-stimulating hormone (FSH)-dependent differentiation of porcine granulosa cells from small follicles for up to six days in culture. Confluent monolayers of epithelioid cells were established after culture on fibronectin-coated culture dishes (FBN, 2 micrograms/cm2) in nutrient medium supplemented with human low-density lipoprotein (LDL, 10 micrograms/ml), insulin (I, 1 microgram/ml), and thrombin (TH, 1 NIH U/ml). Each of these factors was necessary to maintain the epithelioid morphology of the monolayers that attained 70% of the protein content and 71% of the cell number of replicate cultures maintained in nutrient medium supplemented with 10% fetal calf serum and insulin. Addition of FSH to the FBN/LDL/I/TH-supplemented cultures resulted in dose-dependent increases in progesterone secretion and [125I]-iodo-human chorionic gonadotropin (hCG) binding comparable to those obtained in the cultures containing serum. These results indicate that the attachment, epithelioid morphology, and differentiated function of porcine granulosa cells (GCs) can be maintained in defined culture conditions. This culture system will facilitate study of the effects of growth promoters and differentiative agents on GC function in the absence of poorly defined serum supplements.

Full Text

Duke Authors

Cited Authors

  • Buck, PA; Schomberg, DW

Published Date

  • February 1, 1987

Published In

Volume / Issue

  • 36 / 1

Start / End Page

  • 167 - 174

PubMed ID

  • 3105613

Pubmed Central ID

  • 3105613

Electronic International Standard Serial Number (EISSN)

  • 1529-7268

International Standard Serial Number (ISSN)

  • 0006-3363

Digital Object Identifier (DOI)

  • 10.1095/biolreprod36.1.167

Language

  • eng