Faculty Bibliography

Human diploid fibroblast-like cells were maintained in an arrested, essentially non-mitotic state for extended periods of time in culture by lowering the serum concentration in the medium from 10 to 0.5%. These arrested cells re-entered the proliferative state when subcultivated in medium containing 10% serum. The morphological distribution and enzyme activities associated with the Golgi complex were examined during growth, arrest, and recovery. Cells grown in medium containing 10% serum possessed a well-developed Golgi complex consisting of parallel arrays of membranes and associated vesicles. Galactosyl transferase activity was highest after 3 days growth (17.5 +/- 5.0 nmol galactose transferred/45 min/mg protein) and declined to 9.8 +/- 3.0 at day 7. When the serum concentration was reduced to 0.5%, Golgi complexes were rarely observed by electron microscopy and galactosyl transferase activity was further reduced into medium containing 10% serum recovered from the arrested state and ultrastructurally resembled cells continuously cultured at the higher serum level. Numerous Golgi complexes reappeared and galactosyl transferase activity increased to 13.0 +/- 3.34 days after subcultivation. These results indicate that the Golgi complex can be experimentally manipulated in human diploid fibroblast-like cells in a manner which may be useful for the study of the biogenesis of this organelle.

Epithelial-cell enriched primary cultures have been established from rat ventral prostate (RVP). Minced ventral prostates were dissociated with 0.5% collagenase in F12K tissue culture medium containing 1% fetal bovine serum. This treatment resulted in the gradual removal of stromal elements from the base of the epithelial cells. After 60 minutes of digestion the aggregates of epithelial cells were washed and plated at high density in F12K plus 10% horse serum. After 48 hours in vitro the unattached cells were removed from the culture dishes, washed, and reinoculated into new culture vessels containing fresh medium. After 96 hours in vitro, the aggregates had attached to the culture vessels and spread out to yield discrete patches of epithelial cells. By 144 hours in vitro the patches of cells had grown and coalesced to form a semi-confluent monolayer of epithelial cells. Ultrastructrual examination of these cultures indicated that adjacent cells were joined by desmosomes and tight junctions and had formed "lumen-like structures" into which projected microvilli. In addition, the cells contained secretory granules and tonofilaments, giving them a morphological appearance similar to prostate epithelial cells in the intact organ. The primary cultures also retained histochemical activities for acid phosphatase, beta-glucuronidase, and succinic dehydrogenase that were similar to the intact organ.

beta-Histine-HCl has been shown to be effective in modifying the size of developing myocardial infarcts. In the present study the hypothesis that beta-histine increases the flow of blood through collateral channels and thus supplies blood to ligated areas of the myocardium was investigated in the dog. The methods used were measurement of retrograde coronary blood flow and angiography after coronary artery ligation. beta-Histine administration for 6 h increased retrograde blood flow 68.2--91.0% over controls. Coronary angiography demonstrated the existence of collateral channels 200--400 micrometer in diameter within the myocardium after ligation and 4 h of beta-histine administration.