Preparation of an enriched fraction of luteinizing hormone-secreting cells from a mixed population of rat anterior pituitary cells

J. A. Flaws, D. E. Suter

Research output: Contribution to journalArticlepeer-review

Abstract

To determine whether female rat gonadotropes differentially secrete only LH, only FSH, or both LH and FSH, a new method of preparing an enriched fraction of LH-secreting cells from a mixed population of cells was developed. Dispersed female rat pituitary cells were incubated for 4 h in glass tubes with magnetic beads coated with anti-LH antibody. The tubes were then placed in a magnetic particle concentrator to separate cells bound to antibody-coated beads (518b7-bound) from the rest of the cell population (518b7-unbound). Several controls were used in this experiment: 1) mixed cells immediately plated without any preincubation (control); 2) mixed cells incubated without magnetic beads (incubated-only); 3) mixed cells incubated with magnetic beads but not separated (nonseparated); and 4) mixed cells first incubated with magnetic beads coated with normal mouse serum (NMS) and then separated into NMS-bound and NMS-unbound populations. After removal of magnetic beads from cell populations, the cells were plated at 200 000 cells/well. The 518b7-bound cells secreted 1.5-, 2-, 2-, and 5-fold more LH than control, incubated-only, nonseparated, or 518b7-unbound cells, respectively (p < 0.05); 518b7-bound cells secreted 2.5-, 3.8-, 3.8-, and 11- fold more FSH than control, incubated-only, nonseparated, and 518b7-unbound cells. The ratio of LH:FSH secreted did not vary between the different treatment groups, but the ratio of LH:FSH contained in 518b7-selected cells was greater than that for unbound cells. Our data indicate that we have prepared an enriched fraction of LH-secreting pituitary cells that also secrete FSH but have a greater capacity for producing LH than FSH in culture. These results are supportive of the view that LH and FSH are secreted by the same cells.

Original languageEnglish (US)
Pages (from-to)1026-1035
Number of pages10
JournalBiology of reproduction
Volume48
Issue number5
DOIs
StatePublished - 1993
Externally publishedYes

ASJC Scopus subject areas

  • Reproductive Medicine
  • Cell Biology

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