Following adaptation, the ability of the 4 matrices to support iPSC Cell Line 1 and 2 was evaluated. The 4 matrices were used following both the conventional pre-coating protocol, and in-solution to compare the two techniques. ‘In-solution’ involves adding the matrix to the culture media simultaneously with cell passaging, eliminating the need to pre-coat plates.

Method

Cell Line 1 was seeded at 1×10⁴ cells/mL and Cell Line 2 was seeded at 1.3×10⁴ cells/mL into 24 well plates. Wells which had been pre-coated with each of the matrices at their standard coating concentrations were set up in triplicate. Simultaneously, wells which were coated following the in-solution method were set up in triplicate for each matrix. Wells at 0.5x and 0.25x of the standard coating concentration were also set up for each matrix, following both the pre-coating and in-solution method, in triplicate. Finally, control wells, with no matrix present, were set up.

For the pre-coating method, 300µl/well of the appropriate matrix was added and processed as per the matrices’ instructions, before 2mL of cell suspension was added. For the in-solution method, 300µL of the appropriate matrix was added to the 2mL of cell suspension, before aliquoting into the wells.

Plates were cultured for 5 days and assessed on a daily basis for their percentage confluence using the Cell Metric®.