Matrices for Cell Culture – How to develop healthy attachment

Introduction

According to the dictionary, there are many definitions for the word matrix.  From my point of view the following three pertain to cell culture; 1)  a substance in which something is embedded or enclosed; 2) a situation or set of circumstances that allows or encourages the origin, development, or growth of something and; 3) the substance that exists between cells and from which tissue., for example, cartilage and bone, develops.

Many of the common cell types we use are robust and grow on cell culture treated plastic. However, there are many cell types that have difficulty attaching and/or spreading on cell culture treated plastic.  In other cases, cells may attach but not differentiate unless the attachment surface correct. To learn more about matrices or if you are using them and have questions, this Ask The Expert topic is your chance to learn more about attachment surfaces and the rationale for using them.

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Question 1

Can you recommend any matrices that don’t require enzymes during passaging. I am using iPSCs.

This is a good question. You know most of the matrices out there do require some sort of enzymatic digestion if your goal it to obtain single cells. Some matrices are such that the matrix can be dissolved to liberate the cells for assays or passaging. One example is Life Technologies AlgiMatrix 3D where a … Continued

Question 3

Respected Sir, I want to know that, above mentioned Matrices for Cell attachment are autoclavable? What concentration will be effective per square centimeters of surface area?

This is a difficult question to answer since it depends on the matrix you are using. Generally for things like collagen, laminin, fibronectin, poly-L-lysine or something like that, we use 0.1 mg/ml using for example, 2 mls for a 35 mm cell culture dish. I use sterile water to reconstitute and dilute, then I add … Continued

Question 4

Do you know of any available clinical grade quality matrices? Or maybe specialty plates that allow for less or no plate coating for use with iPSCs.

Primorigen Biosciences has something called StemAdhere which they say is a defined matrix for hPSC and I imagine iPSCs. Another is Thermo Scientific Nunclon Vita Stem Cell Culture Surface. This format allows for the direct attachment and growth of cells without the need of feeder layers or matrix coatings. I hope this helps and good … Continued

Question 5

How do matrices affect differentiation? Is there a way to control this?

Yes they do. One of the major reasons people use matrices is to maintain or promote differentiation. Years ago attachment was thought of as simply that…now we know attachment proteins in a cell contacting the proper matrix is a survival signal and a differentiation signal. Many of the different attachment proteins in a cell act … Continued

Question 6

What are the differences, advantages/disadvantages to 3-D matrices?

Lets start with the advantages. When we grow cells the way most people do, by placing them in a dish and allowing them to attach we are growing cells in 2D. As you are aware few cells in the body grow in 2D, they actually grow in 3D. The idea behind 3D culture is to … Continued