Cultivation of human embryonic liver cells in disposable microplates.

Zuckerman, Tsiquaye, and Fulton in 1967 described a method for the cultivation of monolayers of differentiated parenchymal human embryonic liver cells in glass tissue culture plates made especially for this purpose. The apparatus utilized and the precise method of implantation are given in detail by Fulton (1960). This technique was later modified and extended to the culture of very small fragments of human adult liver obtained by needle biopsy for diagnostic purposes (Taylor, Zuckerman, and Farrow, 1970). The present communication describes the adaptation of these methods to the growth of human embryonic liver cells in commercially available disposable plastic microplates and the need to manufacture special glass tissue culture vessels is thus eliminated. This adaptation also facilitates the cultivation of tissue when only very limited amounts are available and renders the method more suitable for wide-scale use by utilizing readily available apparatus. Human embryo livers 8 to 16 weeks old were obtained from the Tissue Bank of the Royal Marsden Hospital and a suspension of cells was prepared by disaggregation of the tissue for 10 minutes in 0 2% trypsin (Zuckerman et al, 1967b). Four cycles of trypsinization were utilized. Dispersed cells were removed after each cycle and kept at 4°C until trypsinization was complete. The trypsin was inactivated with 20% calf serum and a pellet of cells obtained by centrifugation at 700 rpm for five