A modified and flexible procedure is reported for the preparation and cultivation of cells from the normal human amnion. Disinfectants and detergents were found detrimental when used for rinsing the vessels, gloves, and instruments used in collection, and they were avoided. The amnion could be stored at room temperature up to 36 hours after delivery. The intact amniotic membrane was digested with trypsin-buffer solution, and only one extraction was necessary. Petri plate cultures required an atmosphere of approximately 5 per cent CO2. Confluent cultures in petri plates, tubes, and bottles were maintained for several weeks with routine changes of the medium. Although the cells grew in a variety of media, more consistent results were obtained with Parker's 199 + 20 per cent human serum with or without 2 per cent chick embryo extract than with other synthetic media plus human or animal sera.
Aided by the American Cancer Society Institutional Grant 43H.