Characterization of osteoblastic properties of 7F2 and UMR-106 cultures after acclimation to reduced levels of fetal bovine serum.
Estrogen plays an important role in skeletal physiology by maintaining a remodeling balance between the activity of osteoblasts and osteoclasts. In an attempt to decipher the mechanism through which estrogen elicits its action on osteoblasts, experimentation necessitated the development of a culturing environment reduced in estrogenic compounds. The selected medium (OPTI-MEM) is enriched to sustain cultures under reduced fetal bovine serum (FBS) conditions and is devoid of the pH indicator phenol red, a suspected estrogenic agent. This protocol reduced the concentration of FBS supplementation to 0% through successive 24 h incubations with diminishing amounts of total FBS (1%, 0.1%, and 0%). The protocol does not appear to alter the viability, cell morphology, or osteoblast-like phenotype of 7F2 and UMR-106 cell lines when compared with control cells grown in various concentrations of FBS. Although the rate of mitotic divisions declined, the 7F2 and UMR-106 cultures continued to express osteoblast-specific markers and exhibited estrogen responsiveness. These experimental findings demonstrate that the culture protocol developed did not alter the osteoblast nature of the cell lines and provides a model system to study estrogen's antiresorptive role on skeletal turnover.
Ganguly
S
S
Department of Biological and Environmental Science, Morehead State University, 103 Lappin Hall, Morehead, KY 40351, USA.
Ashley
L A
LA
Pendleton
C M
CM
Grey
R D
RD
Howard
G C
GC
Castle
L D
LD
Peyton
D K
DK
Fultz
M E
ME
DeMoss
D L
DL
D000222:Adaptation, Physiological; D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D002417:Cattle; D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004958:Estradiol / Q000494:pharmacology; D005260:Female; D005312:Fetal Blood / Q000502:physiology*; D005819:Genetic Markers; D007150:Immunohistochemistry; D051379:Mice; D010006:Osteoblasts / Q000502:physiology*; D015675:Osteocalcin / Q000096:biosynthesis; D011247:Pregnancy; D053245:RANK Ligand / Q000235:genetics / Q000502:physiology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D044967:Serum / Q000502:physiology*
D005819:Genetic Markers; D053245:RANK Ligand; D015675:Osteocalcin; D004958:Estradiol; D000469:Alkaline Phosphatase