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MethodsPrecoating of culture substratesFor the dose resolution adhesion assay, 96 well flat bottomed number plates (Cell culture medicated; BD Falcon) Were precoated by way of iMatrix 511 (Nippi), Laminin 521 (Biolamina), or even rhVTN N (20 398 aa associated with vitronectin; Wako) At concentrations of 0 4g/cm2 for 3h at 37C in a CO2 incubator just prior to cell seeding. For adhesion efficiency assays and passaging to compare with the uncoated manner, 24 or 6 well plates (BD Falcon) Were layered with iMatrix 511 at 1g/cm2. Dulbecco’s phosphate buffered saline (DPBS) was applied to dissolve the culture substrates. the answer for any volume for coating was standardised to 100l/cm2, But altered to 50l/well for 96 well plates.
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