Trypsin 1 mm edta solution

Four independent cell lines consisting of different ratios of haploid and diploid cells [from 20% to 70% haESC, Fig. 2(a)] were thawed from cryostorage and cultured for up to 10 passages. Cell cultures were split and passaged every 2 days either using the separation device or without. Briefly, cells were washed twice with phosphate-buffered saline (PBS, pH 7.4; Life Technologies) for 2 min and incubated with a 0.25% trypsin/1 mM EDTA solution (Thermo Fisher) for 4 min at 37 °C. A single cell suspension was prepared by mechanical rocking of the plate, inactivation of trypsin by adding the serum containing medium, and pipetting several times before centrifugation at 1000 m s⁻². The supernatant was discarded, and the cells were resuspended in 2 ml serum and LIF media. 0.5 ml aliquots were analyzed to assess the haploid cell content by flow cytometry. An aliquot of the cell suspension was then plated into a new dish either without purification or using the separation unit (n = 3 for each treatment).

PDAC cell lines (MIA PaCa-2 and PANC-1) were purchased from the American Type Culture Collection and grown at 37°C with 5% CO₂. MIA PaCa-2 cells were cultured in DMEM high-glucose medium (Thermo Fisher Scientific) supplemented with 10% fetal bovine serum (Atlanta Biologicals) and 2.5% horse serum (Corning). PANC-1 cells were cultured in DMEM high-glucose medium supplemented with 10% fetal bovine serum. The cell lines were sub-cultured by enzymatic digestion with 0.25% trypsin/1 mM EDTA solution (Thermo Fisher Scientific) when they reached approximately 70% confluency.