PP-027 - Maturing the oocyte: A comparison of maturation media and the effect of fetal calf serum supplementation

An aliquot from a continuous fermenter culture of baby hamster kidney cells (BHK-21 Clone PD-4) (Wistar) maintained in Ca free Eagle's Basal Medium containing 2% Kaolin adsorbed fetal calf serum was planted in spinner flasks at 300,000 cells per ml, total volume 600 ml. After equilibration for one day at 35°C to insure that cells were in log phase, the culture was infected with the M-33-AGMK25 BHK-219 strain of rubella at an input multiplicity of about 6 TCID50 per cell. The virus was identified with specific rubella antiserum.Preliminary experiments had shown that such cultures would reach a peak or plateau HA titer of approximately 1:64, 24 hrs after inoculation and would continue to yield virus for 6 to 12 days. One hundred ml aliquot harvests were withdrawn daily and the culture was returned to volume with growth medium and incubation continued. The harvested cells were spun down rapidly at 2500 rpm per 15 mins., fixed in 3.7% gluteraldehyde in Ca free phosphate buffer saline, and post fixed in osmium tetraoxide. After dehydration, the cells were embedded in Epon 812 and cured approximately 20 hrs at 60°C.

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Fish kidney cell line in response to heat shock

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100123921 ◽

1992 ◽

Vol 50 (1) ◽

pp. 704-705

Author(s):

Li-Chu Tung ◽

Yung-Reui Chen ◽

Shiu-Nan Chen ◽

Guang-Hsiung Kuo

Keyword(s):

Heat Shock ◽

Cell Line ◽

Fetal Calf Serum ◽

Calf Serum ◽

Uranyl Acetate ◽

Ultrastructural Changes ◽

Heat Shock Treatment ◽

Acanthopagrus Schlegeli ◽

Cacodylate Buffer ◽

Fish Kidney

In the present study, the ultrastructural changes of BPK cells, a fibroblast-like cell line, derived from the kidney of juvenile black porgy Acanthopagrus schlegeli, under heat shock treatment are described.The BPK cells were maintained in L-15 medium supplemented with 10% fetal calf serum and 0.15 M NaCl at 28|C2. The heating was carried out in precalibrated water baths. Monolayers of cells, grown on coverslips in parafilm-sealed petri dishes were submerged under water for 30 min at 40|C treatments. Cells were fixed in 2.5% glutaraldehyde in 0.1 M cacodylate buffer supplemented with 6.6% sucrose, postfixed in 1% OsO4 and flat embedded in Spurr’s resin. Silver section were cut parallel to the substratum, stained with uranyl acetate and Reynold’s lead citrate, and examined in a Hitachi H-600 electron microscope at 75 KV.

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Fetal calf serum contains activities that induce fetal hemoglobin in adult erythroid cell cultures

Blood ◽

10.1182/blood.v75.9.1862.1862 ◽

1990 ◽

Vol 75 (9) ◽

pp. 1862-1869 ◽

Cited By ~ 13

Author(s):

P Constantoulakis ◽

B Nakamoto ◽

T Papayannopoulou ◽

G Stamatoyannopoulos

Keyword(s):

Cell Cultures ◽

Fetal Calf Serum ◽

Calf Serum ◽

Fetal Hemoglobin ◽

Erythroid Cell ◽

Erythroid Progenitors ◽

Globin Mrna ◽

Erythroid Progenitor ◽

Fetal Sheep ◽

Gamma Globin

Abstract Cultures of peripheral blood or bone marrow erythroid progenitors display stimulated production of fetal hemoglobin. We investigated whether this stimulation is due to factors contained in the sera of the culture medium. Comparisons of gamma/gamma + beta biosynthetic ratios in erythroid colonies grown in fetal calf serum (FCS) or in charcoal treated FCS (C-FCS) showed that FCS-grown cells had significantly higher gamma/gamma + beta ratios. This increase in globin chain biosynthesis was reflected by an increase in relative amounts of steady- state gamma-globin mRNA. In contrast to its effect on adult cells, FCS failed to influence gamma-chain synthesis in fetal burst forming units- erythroid (BFU-E) colonies. There was a high correlation of gamma- globin expression in paired cultures done with C-FCS or fetal sheep serum. Dose-response experiments showed that the induction of gamma- globin expression is dependent on the concentration of FCS. These results indicate that FCS contains an activity that induces gamma- globin expression in adult erythroid progenitor cell cultures.

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