The response of human peripheral blood lymphocytes to phytohemagglutinin: determination of cell numbers
| Autor: | Stewart F. Cramer, Carleton C. Stewart, Palmer G. Steward |
|---|---|
| Rok vydání: | 1975 |
| Předmět: |
Lysis
Lymphocyte Iron Immunology Cell Cetrimide Blood Sedimentation Cell Separation Biology Lymphocyte Activation Tritium Andrology Leukocyte Count Lectins medicine Doubling time Humans Lymphocytes Cells Cultured Cell Nucleus Staining and Labeling DNA In vitro Peripheral blood Kinetics medicine.anatomical_structure Volume (thermodynamics) Pronase Erythrocyte Count Autoradiography Cell Division Thymidine |
| Zdroj: | Cellular immunology. 16(2) |
| ISSN: | 0008-8749 |
| Popis: | Optimization and definition of conditions for studying lymphocyte function in vitro resulted in exponential proliferation of lymphocytes from day 2 to day 5 with an average doubling time of 20 hr. The number of cells in culture on day 5 was 5–10 times as great as the number initially planted and 10–20 times as great as the number surviving in culture on day 2. An improved pronase-cetrimide technique was used to determine the number of viable lymphocytes as a function of time after addition of PHA. The volume changes in nuclei, obtained after cetrimide treatment, were quantitated using a curve-fitting computer program. The response could be described in terms of an induction phase (0–2 days) characterized by a decrease in cellularity and an increase in nuclear volume, a proliferation phase (2–5 days) characterized by an exponential proliferation and a continued increase in the number of cells having a large nuclear volume, and a lysis phase (5–14 days) characterized by a decrease in cellularity and a decrease in nuclear volume. The results reported here suggest that the ratio of the number of cells cultured to the volume of culture medium was crucial for optimal transformation and proliferation, 105 cells/ml producing far better responses than 106 cells/ml. |
| Databáze: | OpenAIRE |
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