Observation of production of immunoactive prolactin by normal human connective tissue in cell culture.
Data from our in vitro studies indicate a new source of prolactin (PRL)-like activity, normal human connective tissue. Fascial cells from primary culture and subsequent passages produced an extracellular antigen which specifically reacted in a radioimmunoassay RIA developed to detect human pituitary PRL. An initial peak or first surge of fascial PRL-like activity occurred between 4 and 15 d in primary culture. Ibuprofen, cytotoxic levels of 0.01% azide, or 7.5 mM EDTA and medium lacking serum [fetal bovine serum (FBS)] significantly (P less than or equal to 0.05) reduced PRL-like activity levels, whereas female steroids, 257 to 342 milliosmolarity, 1 to 3.6 mg/ml glucose, 2 to 20% FBS, and dialyzed FBS (MWCO approximately 1 kDa) were without effect. Optimum production of PRL-like activity occurred at pH 7.3. A second surge began after 18 d and continued until passage indicating that perhaps two populations of cells produced PRL-like activity in primary culture. Production of PRL-like activity by cells from early passages (1 and 2) became detectable at confluence, was serum-dependent, showed two patterns (tonic, rising to plateau), and averaged 3.2 fg.cell-1.3 d-1 feed interval. Cells from late passages showed morphologic damage from repetitive trypsinization, aging, and reduced production of PRL-like activity with aberrant production pattern. Production of PRL-like activity was maintained in an unusual long-term culture. These in vitro studies demonstrate the most recently recognized and ubiquitous source of human extrapituitary PRL or PRL-like activity, normal connective tissue (fascia).
Chapitis
J
J
Department of Obstetrics and Gynecology, University of Vermont College of Medicine, Burlington 05405.
Betz
L M
LM
Brumsted
J R
JR
Gibson
M
M
Kuslis
S T
ST
George-McDaniel
H
H
Riddick
D H
DH
D000009:Abdominal Muscles; D001386:Azides / Q000494:pharmacology; D001769:Blood; D002478:Cells, Cultured; D003238:Connective Tissue / Q000187:drug effects / Q000378:metabolism*; D003470:Culture Media; D004492:Edetic Acid / Q000494:pharmacology; D004958:Estradiol / Q000494:pharmacology; D005947:Glucose / Q000494:pharmacology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007052:Ibuprofen / Q000494:pharmacology; D007700:Kinetics; D009994:Osmolar Concentration; D011374:Progesterone / Q000494:pharmacology; D011388:Prolactin / Q000096:biosynthesis*; D011863:Radioimmunoassay
D001386:Azides; D003470:Culture Media; D011374:Progesterone; D004958:Estradiol; D011388:Prolactin; D004492:Edetic Acid; D005947:Glucose; D007052:Ibuprofen