We have recently developed new techniques for culturing chronic lymphocytic leukemia (CLL) lymphocytes from human aqueous humor. We used leukemic lymphocytes collected from the aqueous humor of a patient with CLL and leukemic glaucoma. We grew these leukemic cells in combination with a feeder cell layer and other technical refinements. Microscopy and immunoassay indicate success in obtaining a homogenous population of B-type CLL cells through the 12th passage of the culture. No significant effect on cell growth was found with either of two mitogens (PWM and PHA), or between culture with and without autologous serum. Our new techniques for culturing leukemic cells derived from the aqueous humor provide a reliable resource for the study of chronic lymphocytic leukemia lymphocytes and leukemic glaucoma.