Cancer cell isolation kit c100xx

Four to six week-old female Syrian Hamsters were anesthetized using 3% isoflurane; their cheek pouches were painted with 0.5% DMBA (9, 10-dimethyl-1, 2-benzanthracene #D3254,Sigma) in paraffin oil (#18512, Sigma) using camel hair brush No.4 (#6020-04000, Gordon Brush) twice a week for 12 to 14 weeks ^{24 (link), 25 (link)}. The tumors were aseptically harvested from euthanized animals and transferred to RPMI media without serum. The cells from the tumors were prepared using a commercial kit (Cancer cell isolation kit - #C100XX, Panomics, Affymetrix) and plated in DMEM supplemented with 20% FBS, 10 μg/mL of hydrocortisone, 50 ng/mL of epidermal growth factor (EGF), 5 μg/mL of spermine and antibiotics ^{26 (link)}. The cells were further cultured in DMEM supplemented with 10% FBS with antibiotics. DMBA-induced buccal tumor cells (2 × 10⁷⁾ were transplanted subcutaneously in the flanks. Ten days after injection the tumors were aseptically removed and the cells were prepared and clonal cell lines (HPT11 and 12) were established.