The simultaneous use of two prostaglandin radioimmunoassays employing two antisera of differing specificity. II. Relative stability of prostaglandins E1, E2 and F1alpha in cell cultures of BALB/c 3T3 and SV3T3 mouse fibroblasts.
The relative stability of Prostaglandins (PGs) E1, E2 and F1alpha in cultures of BALB/c 3T3 and SV3T3 cells has been evaluated using 3 different approaches. First, total recovery of tritium in the ethyl acetate phase following incubation and extraction of PGF1alpha and PGE1 demonstrated greater stability for PGF1alpha (88.8%) than PGE1 (65.9%). Second, analysis of incubated, extracted, tritiated PGs by thin layer chromatography revealed decreases of up to 23% in the PGE zone following incubation of 3H-PGE1. With increasing time of incubation, decreases in the PGE zone were accompanied by increase in PGA-like compounds. 3H-PGF1alpha demonstrated greater stability, having greater than 90% recovery of the tritium in the PGF zone. A third approach to the assessment of PG stability in culture was the comparison of the production of individual PGs by radioimmunoassay (RIA). The data obtained by RIA indicated a lag in the increase of PGA and PGB, until an initial rise in PGE was noted, suggesting that PGA and PGB may be secondary products arising from PGE which exhibits only partial stability in culture. By employing two RIAs, one for total PGE and one for PGA and PGB, the composite determination PG [E + (A + B)] can be used to provide a more meaningful determination of PG production because of the instability of the PGs. On the other hand, individual determinations are helpful in assessing the stability of PGEs in cell cultures.
D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D002855:Chromatography, Thin Layer; D005347:Fibroblasts / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D011458:Prostaglandins E / Q000032:analysis / Q000378:metabolism*; D011460:Prostaglandins F / Q000032:analysis / Q000378:metabolism*; D011863:Radioimmunoassay; D013997:Time Factors; D014316:Tritium
D011458:Prostaglandins E; D011460:Prostaglandins F; D014316:Tritium