Autoradiographic Studies of Human Lymphocytes Cultured in Vivo

Abstract

Cell culture studies with phytohemagglutinin (PHA) have clearly established the small lymphocyte as a cell capable of reverting from a quiescent state to a blast form that synthesizes DNA and undergoes division. The relationship of this in vitro proliferation to in vivo function, remains to be elucidated. However, by its resmblance to the behavior of lymphocytes stimulated immunologically in vivo ( l ) , the in vitro response to PHA may represent a model of the cell's immunologic competence. The present paper reports on the proliferation of human lymphocytes cultured in diffusion chambers placed into the peritoneal cavities of rats. This system has yielded consistently good results when applied to rat lymphocytes which are difficult to maintain in vitro (2-5). The diffusion chamber technique may thus represent an improvement in the method of culturing lymphocytes, as it more closely approximates the true in vivo environment.Materials and Methods. The technique for culturing blood lymphocytes in diffusion chambers was previously described (2). Heparinized blood (100 units/ml) obtained from a healthy donor, was centrifuged at 6513 for 8 min and the leukocyte-rich supernatant was removed. Phytohemagglutinin-P (Difco PHA-P, reconstituted in 30 ml of medium 199) was added to one half of the leukocyte suspension at a concentration of 0.2 ml/ 6 X

Cytogenetics and Fanconi's Anemia: experimental and other studies of a family

“…The cell cycle of human lymphocytes ln_ yltro and in vivo has been investigated by others [4,10,21,61,122,131], and the uncorrected normal cell cycle times given here agree well with the raw data provided in those reports. The corrected normal labelling indices from PLCp curves agree well with those found by MacKinney «st, aX, but their mitotic indices were lower than those found in this study [71].…”

Return of Alkaline Phosphatase in Chronic Myelocytic Leukemia Cells in Diffusion Chamber Cultures

“…Kinetics of proliferation of normal human bone marrow ( B M ) and peripheral blood cells have been studied in the diffusion chamber culture system (1)(2)(3)(4). I n this preliminary report the growth pattern, cell survival kinetics, leukocyte alkaline phosphatase (LAP) activity changes, and chromosomal analysis of two chronic myelocytic leukemia (CML) patients' BhI cells cultured in the chamber system are presented.…”

Temporal Variation in Mitotic Index of Phytohaemagglutinin Stimulated Human Lymphocytes

“…It causes synthesis of DNA and the onset of mitosis in a proportion of cells [9,10] which have been identified as almost all being blast cells [12]. Hence, since transformation is relatively constant within an individual it is possible that DNA synthesis, which is necessary for mitosis to occur, is also relatively constant.…”