Welcome to our reagent blog!
For this month’s reagent round up, I would like to share a comparative data set showcasing our MatriClone matrix.
MatriClone is an exciting opportunity for customers looking to accelerate their cell therapy workflows and deliver new treatments to market. For those who are not familiar with the matrix, it is comprised of truncated recombinant Laminin-511 protein and is designed to support the health and pluripotency of induced pluripotent stem cells (iPSCs). Unlike most iPSC matrices, MatriClone is optically suited for whole well imaging required for regulatory approval and is soluble, meaning that pre-coating of plates is no longer required!
Introduction
Two iPSC cell lines, Cell Line 1, and Cell Line 2 were adapted to 4 iPSC matrices. These included MatriClone and 3 other iPSC 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 the in-solution method to compare the two techniques. For those who are not familiar with the ‘in-solution’ method, this involves adding matrix to the culture media at the same time as the cells during cell passaging, rather than pre-coating the plate.
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 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, containing no coating, were also set up.
For the pre-coating method, 300ul/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, 300uL 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 and subsequent growth were observed when Reagents B and C were used in-solution, by comparison.
The next observation of note is that 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, both cell lines did not attach and grow in the absence of matrices.
Conclusion
The study demonstrates MatriClone as a robust matrix to support the health and growth of iPSCs. Comprised of truncated recombinant Laminin-511 protein, the matrix promotes greater adhesion than full-length Laminin protein and is optically suited for automated imaging. Finally, with the additional advantage of being suitable for in-solution applications, MatriClone is an exciting opportunity for customers to accelerate their cell therapy workflow and deliver new treatments to market.
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 the research and GMP iPSC workflows. Clinical Manufacturing Grade Laminin is available for use with VIPS® PRO, VIPS® , Cell Metric® X or Cell Metric®.
I hope you have enjoyed this short case study. Please reach out to us at [email protected] if you would like to learn more about MatriClone and Clinical Manufacturing Grade Laminin!
