P-069

Matrigel replacement: Evaluation and development of alternative cell culture coatings for induced pluripotent stem cells

PD Dr. Anja Wilmes,
Vrije Universiteit Amsterdam

01/2020-06/2021

Matrigel is used to culture stem cells in the laboratory. For its production mice have to be induced with tumours and sacrificed. New animal-free products could be a better alternative.

Human cell culture systems play an important role in research, for example in toxicology, where they are used to understand the mechanisms of actions of medicines or toxins. In particular so called human induced pluripotent stem cells (iPSC) are expected to be of great importance in the development of more relevant, animal-free test systems. iPSC can be generated from adult cells, e.g., blood cells, and in theory can be differentiated into any cell type of the body. In our laboratory, we have differentiated iPSC into different cells of the kidney, including podocytes, to test compounds for renal toxicity.

However, for culturing iPSC, a coating made of a specific extracellular matrix is necessary that allows the cells to attach to the culture dish. For this, a product called Matrigel is widely used. Matrigel is an animal product and for its production mice have to be induced with sarcoma tumours. The weight of these tumours often reaches up to 20% of total body weight of the mice before they are finally sacrificed.

While Matrigel is one of the most commonly used coatings for culturing iPSC, its replacement is desired for several reasons, including better standardization conditions and of course for ethical reasons. Even though some alternative products are commercially available, these are only used sparely in laboratories. Reasons for this vary and include often higher costs of alternative products and limited independent studies that compare different products from different companies.

In this project one of our aims is to carefully evaluate and compare existing products, with a major focus on iPSC quality and stability: iPSC have to remain pluripotent and should not acquire genetic variations. Furthermore, we are working on the development of new and more cost-effective coatings by employing human cell lines.

The third aim of this project is to test these coatings for the differentiation of iPSC into renal cell types (e.g. podocytes) in order to develop a new differentiation protocol that is independent from Matrigel. Here, we will analyse podocyte specific markers, including synaptopodin and WT1.

 

Fig. 1:   iPSC-derived podocytes are growing on either Matrigel (A) or on different Matrigel-free coatings (B-D).

 

 

Institution

Division of Molecular and Computational Toxicology
Vrije Universiteit Amsterdam,
De Boelelaan, 1108,
1081 HZ Amsterdam, The Netherlands.

Duration

01/2020 - 06/2021