Preparation and culture of dispersed avian pituitary cells, and age-related changes in donor pituitary weight and growth hormone content.
Techniques were perfected for the enzymatic dissociation of chicken pituitary glands and a number of factors evaluated for their effects upon growth hormone (GH) production by dispersed chicken pituitary cells in culture. Age-related changes in donor pituitary weight and GH content were also determined. A procedure involving digestion of minced glands with a solution of 0.1% trypsin in S-MEM tissue culture medium (0.1% BSA) for 1 hr at 37 degrees under an atmosphere of 5% CO2-95% air yielded greater than 2.0 X 10(6) cells per gland with 80-90% viability. Five tissue culture media (D-MEM, alpha-MEM, RPMI 1640, Med-199, Earle's salts), two serum sources (calf serum (CS), horse serum (HS), and two levels of serum (5, 20%) were tested for their ability to support GH synthesis over 4 days in culture. Additionally, two culture regimes (continuous culture vs daily media changes) were evaluated for their effects on GH production. alpha-MEM resulted in the numerically highest net GH synthesis (over starting cell content), although not statistically different from RPMI 1640 or Earle's salts. Neither serum type nor percentage was significant; therefore the lower serum percentage (5) was adopted for future studies. Culture regime significantly altered the proportion of secreted vs stored hormone harvested at the end of the culture period. Changing media daily resulted in a 40% reduction in final cell GH content compared to continuous culture, whereas total cumulative media GH was approximately 39% greater (P less than 0.01). Pituitary weight increased with age until approximately 9 weeks, whereas GH content plateaued earlier, at 5 weeks of age.(ABSTRACT TRUNCATED AT 250 WORDS)
D000375:Aging / Q000502:physiology*; D000818:Animals; D002478:Cells, Cultured; D002645:Chickens; D005260:Female; D013006:Growth Hormone / Q000378:metabolism*; D009929:Organ Size; D010902:Pituitary Gland / Q000166:cytology / Q000502:physiology*; D011863:Radioimmunoassay
D013006:Growth Hormone