Cell Line Development Knowledge Center

Exploring iPSC Matrices: A Comparative Study Exploring the Application of MatriClone­™

MatriClone is a xeno-free matrix which supports the health and pluripotency of induced pluripotent stem cells (iPSCs). Unlike most iPSC matrices, MatriClone is optically suited for imaged-based clonality assurance, and is soluble, meaning that pre-coating of plates is no longer required. In this study, we evaluate MatriClone’s ability, versus 3 other commercially available matrices, to support the growth of iPSCs within 24 well plates. We find that MatriClone facilitates superior adhesion compared to the market alternatives considered within the study, resulting in increased colony growth rates.

Two iPSC cell lines; Cell Line 1, and Cell Line 2, were adapted to 4 different iPSC matrices. These included MatriClone and 3 other matrices which have been anonymized as follows: Reagent A, Reagent B, and Reagent C.

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®.

Results

Cell Line 1 and 2 showed little difference in growth rates when MatriClone was used to pre-coat plates, versus in-solution. There was also minimal difference in growth rates between the different concentrations tested for both methods.

Meanwhile, both cell lines adhered and formed colonies when MatriClone and Reagent A were used in-solution. No attachment or growth was observed when Reagents B and C were used in-solution, by comparison.

Cell Line 1 and 2 both attached and grew at all matrix concentrations when the conventional pre-coating method was followed, however the growth of both was noticeably slower when Reagent C was used.

Finally, neither cell line adhered or grew in the absence of a matrix.

Conclusion

The study demonstrates MatriClone as a robust matrix to support the health and growth of iPSCs. Comprised of the recombinant Laminin-511 E8 fragment, MatriClone promotes greater adhesion than full-length Laminin proteins. While the other matrices did facilitate iPSC growth, there were limitations around their application.

Advanced Instruments are uniquely positioned to provide both research grade MatriClone, and Clinical Manufacturing Grade Laminin from the same source material, ensuring quality and consistency across research and GMP iPSC workflows*.

Thank you for joining us to review this case study. Please reach out to us at [email protected] if you would like to learn more about MatriClone and Clinical Manufacturing Grade Laminin!

Contact us to learn which is right for you!

*Clinical Manufacturing Grade Laminin is available for use with Solentim instrumentation.

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