An in vitro microassay for lymphotoxin using microculture plates and the multiple automated sample harvester
| Autor: | Kenneth W. Sell, Aftab Ahmed, R C Knudsen |
|---|---|
| Rok vydání: | 1974 |
| Předmět: |
Microculture
Chromatography Cell growth Lymphoblast Immunology Liquid scintillation counting Fibroblasts Biology Lymphocyte Activation Tritium Molecular biology In vitro Culture Media Automation Lymphotoxin Cell culture Methods Immunology and Allergy Lymphocytes Lymphotoxin-alpha Cells Cultured Thymidine |
| Zdroj: | Journal of Immunological Methods. 5:55-63 |
| ISSN: | 0022-1759 |
| DOI: | 10.1016/0022-1759(74)90045-3 |
| Popis: | An in vitro microassat for lymphotoxin (LT) using mouse L-929 fibroblasts as target cells grown in the wells of microculture plates is described. The microassay uses 1000 targets cells and 50 μl of LT-containing fluids per well. Destruction or inhibition of target cell growth in the presence of LT-containing fluids is measured by their ability to incorporated 3 H-methyl-thymidine. Radioactively labelled cells are harvested onto glass fiber filter discs using a multiple automated sample harvester and are subsequently counted in a liquid scintillation spectrometer. Inhibitory or destructive activity of LT-containing fluids can be conveniently represented as the dilution which inhibits target cell growth by 50 percent (ID 50 ). The advantages and limitations of the microassay are shown using LT-containing supernatant fluids from human lymphoblast cell lines and PHA-P- and PHA-M-stimulated human lymphocytes. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|