Temporary remissions in acute leukemia in children produced by folic acid antagonist 4-aminopteroylglutamic acid (aminopterin
Scientists at the New York State Animal Health Diagnostic Center at Cornell University and at the New York State Food Laboratory tested three cat food samples provided by the manufacturer and found aminopterin
in two of them.
Attempted abortion with aminopterin
The fused cells were cultured in 96-well plates in DMEM (Invitrogen) containing 200 mL/L fetal calf serum, 200 mmol/L glutamine, 10 g/L OPI (oxaloacetic acid, pyruvic acid, insulin), and 20 g/L HAT (hypoxanthine, aminopterin
, thymidine; Sigma) at 37 [degrees]C in a 5% C[O.sub.2] atmosphere for 10-14 days.
After several minutes the suspension was again centrifuged to pellet the cells, and the PEGM + RPMI medium was removed and replaced with RPMI supplemented with BSA (15% bovine serum albumin), OPI (0.15% oxaloacetate, 0.05% pyruvate, 0.0082% bovine insulin), and HAT (5 X [l0.sup.-5] M hypoxanthine, 2 X [10.sup.-7] M aminopterin
, 8 X [10.sup.
Hybridomas were grown in DMEM culture medium (Sigma) containing 20% fetal calf serum (FCS) (Seromed, Germany), penicillin (100 IU/ml) and streptomycin (100 [micro]g/ml) and supplemented with hypoxantine (1 x [10.sup.-4] M), aminopterin
(4 x [10.sup.-7] M) and thymidine (1.6 x [10.sup.-5] M) (HAT) (Sigma).
Hybrid cells were cultured in medium containing aminopterin
, hypoxanthine, and thymidine (HAT medium) with 100 mL/L fetal calf serum.
Optimem 1 with Glutamax-1 and HAT supplement (hypoxanthine, aminopterin
, and thymidine) were products of Life Technologies.
The fused cells were cultured with 96-well culture plates (Nunc) in RPMI 1640 (Nissui) containing fetal bovine serum, hypoxanthine, aminopterin
, and thymidine.
Cells were cultured at 37[degrees]C and 5% C[O.sub.2] in DMEM (Eurobio) supplemented with 100 mL/L heat-inactivated fetal calf serum (FCS; BioWhittaker), 2 mmol/L glutamine, and 50 mL/L hypoxanthine aminopterin
thymidine (FCS medium).
Production of functional human T-T hybridomasin selection medium lacking aminopterin
Splenocytes were fused with a nonsecretor cell line sp2/0 and plated into Dulbecco's modified Eagle's medium containing hypoxanthine, aminopterin
, and thymidine with 150 mL/L fetal bovine serum.