Evaluation of metabolic pathway activity in cultured skin fibroblasts and blood leukocytes

Analysis of genetic complementation by whole-cell microtechniques in fibroblast heterokaryons.

“…The procedure for detecting ['4C]propionate incorporation in cells in situ by using an autoradiographic method was described by Hill and Goodman (18). It has also been adapted to the direct determination of radioactivity by scintillation counting (19,20). Several modifications were instituted in order to apply it to the microtechnique.…”

“…The day after plating, the cells were incubated at 37"C in the presence of the [14C]propionate medium. After up to [16][17][18][19][20] hr, the reaction was stopped by washing twice with ice-cold phosphate-buffered saline (21), and once, for 10 min, with ice-cold 10% trichloroacetic acid. The cells were rinsed thoroughly with water and were removed by exposing them to 0.3 ml of a 1:10 dilution of concentrated ammonium hydroxide in water until they were completely lysed as observed by microscopy.…”

In vivo andin vitro responsiveness to vitamin B12 in a case of methylmalonic aciduria

“…Her weight has increased steadily and development has improved significantly over the first year of treatment. (Rozen et al, 1977). The cells were washed several times in 10% (w/v) trichloroacetic acid and digested in Soluene-350 (Packard Instruments Ltd) prior to incorporation into scintillation fluid.…”