Back to Search
Start Over
Analyses for in vitro growth conditions, cytokinetics, and sister chromatid exchanges in lymphocytes cultured from the Sprague-Dawley rat
- Source :
- In vitro cellulardevelopmental biology : journal of the Tissue Culture Association. 21(10)
- Publication Year :
- 1985
-
Abstract
- Peripheral blood samples from Sprague-Dawley rats gave successful lymphocyte growth in GIBCO: 1A, RPMI 1640, and Eagle's minimum essential medium (MEM) culture media. Various growth conditions, cytokinetics, and sister chromatic exchange (SCE) induction were studied using reconstituted GIBCO 1A only. Neither methoxyflurane anesthesia of the rats before sampling nor washing of the cells with phosphate buffered saline affected the mitotic index. Cultures treated with [3H]thymidine showed the lymphocytes entering into DNA synthesis after approximately 24 h. The time at which BUdR (5-bromo-2' deoxyuridine) was added, i.e. 0 vs. 24 h incubation, had minimal effect on the mitotic index of cultures harvested at 48 h. However, when harvest was extended to 72 h, mitotic activity was greater in the cultures treated with BUdR at 24 h. No significant differences in mitotic index and the number of average lymphocyte division were detected in cultures exposed to 0.3 to 0.5 microgram/ml BUdR at 24 h and harvested at 72 h. Although SCE frequencies increased in the presence of BUdR, the baseline level of SCEs was estimated to be 5 to 6/cell. Average generation time of the lymphocytes dividing between 48 and 72 h was 16.5 h. Because of its simplicity of culture and the reproducible nature of its in vitro growth kinetics, the Sprague-Dawley rat lymphocyte is a suitable model for cytogenetic investigations.
- Subjects :
- Male
Mitotic index
Lymphocyte
Clinical Biochemistry
Sister chromatid exchange
Plant Science
Biology
chemistry.chemical_compound
medicine
Mitotic Index
Sister chromatids
Animals
Lymphocytes
Incubation
Cells, Cultured
Cell Cycle
Rats, Inbred Strains
Cell Biology
DNA
Molecular biology
Deoxyuridine
Culture Media
Rats
medicine.anatomical_structure
chemistry
Bromodeoxyuridine
Cell culture
Karyotyping
Immunology
Thymidine
Anesthesia, Inhalation
Sister Chromatid Exchange
Biotechnology
Developmental Biology
Subjects
Details
- ISSN :
- 08838364
- Volume :
- 21
- Issue :
- 10
- Database :
- OpenAIRE
- Journal :
- In vitro cellulardevelopmental biology : journal of the Tissue Culture Association
- Accession number :
- edsair.doi.dedup.....f17c6b643ac2510ce88f8f30aa04e56e