Use Of Plasma-Free Medium For In-Vitro Culture Of Human Lymphocytes

A modification of the plasma-free medium of Coulson has been used to study the response of human lymphocytes, cultured in vitro, to phytohemagglutinin (P.H.A.). It has been found that reproducible and satisfactory transformation can be obtained in such a medium, comparable to that found in cells cultured in plasma-containing medium. A low dose of P.H.A., a cell population of 2 × 106 cells per culture tube, and the inclusion of a platelet-associated material are needed to produce successful cultures. The enhancement of P.H.A.-induced transformation of lymphocytes cultured in medium containing plasma by the addition of low molar concentrations of hydrocortisone, methotrexate, and arsenic trichloride is shown. Such stimulation is not seen in plasma-free medium cultures, suggesting that plasma components are essential for such stimulation and excluding synchronization of the cells as a cause for such enhanced transformation in the case of methotrexate. The plasma-free medium has been used to study the reactivity of cancer patients' lymphocytes. Three groups can be recognized: 1. those with comparable function compared to control subjects; 2. those with striking depression when cultured in medium containing autologous plasma, and 3. those with an intrinsic defect in lymphocyte response to P.H.A.