Good question. One time I coated inserts with Col-II originated from Chicken for human stem cells. It worked very well. Since someone has used it than I tried and it worked. Someone asked me this question why not to use human Col II. In my opinion, it will work in your case, you can use any species. Try and let us know.

Hello Nghia Thi Pham

Extracellular Matrix (ECM) proteins coating have proven to be efficient methods to better mimic in vivo cell behavior. The components of ECM are categorized into proteoglycans and fibrous proteins (Collagen, fibronectin and laminin). These components give structural support and facilitate cellular communication.

Depending on the cell type and the area of research you may decide on the ECM proteins.

Collagen is the most abundant protein in mammals. In cell culture it is used to study growth, differentiation, and migration of cells. You may use Collagen Type I for fibroblasts or endothelial cells. Collagen Type II may be used for Chondrocytes and epithelial cells.

Fibronectin is a multifunctional protein involved in cell adhesion and spreading. It also regulates cellular morphology, cell migration, cytoskeletal organization, hemostasis and wound repair. For cell types such as epithelial cells, mesenchymal cells, neuronal cells, fibroblasts cells and endothelial cells you may use fibronectin.

Laminin acts as glue, which holds cells and ECM together. It has active domains for collagen binding, cell adhesion, heparin binding and neurite outgrowth fragment. Laminin modulates cell growth, motility and signaling pathways. Cell types used are epithelial cells, endothelial cells, muscle cells, schwannoma and hepatocytes.

Vitronectin promotes cell migration, proliferation, differentiation and spreading of endothelial and neoplastic cells. The cell types which may be used are endothelial cells, stem cells, melanoma and osteosarcoma.

It is not necessary that the origin of ECM proteins should match the cell line species, but the type of cells you are working with and the area of research does matter in selecting the ECM proteins to coat the cell culture dishes.

Best.

Hi Nghia Thi Pham

If I could add something more after the answer above...

I don't know the area of your research, however, as you work with human cell lines, remember that regarding different origin of ECM proteins, for example laminin made from mouse sarcoma cells, the clinical application is forbidden. Also, the use of recombinant/human proteins (surely laminin) is very expensive. Tell us what you decide. ;)

Best!

You are right also since you need to get approved from ethics board. Than try to use human ECM protein for human, I believe. My research area was to try human stem cell-derived cartilages to treat induced osteoarthritis in nude rat. Yes, If my project goes to human to human one day, than there will be issue, than I will use human collagen II coating for human stem cells.

Nghia Thi,

I assume that you intend to acquire your different ECM preps from commercial suppliers. Do not forget that these are very likely to be purified preparations and highly unlikely to be pure. If run a simple SDS-PAGE you will get some idea of just how impure these preps are!

So what?

Your question suggests that you looking to assay cellular responses to different preps. My question (warning) is how will you resolve responses to majority ECM species (Col, Lam, etc), from responses to minority/trace contaminants? Many contaminants, including breakdown products, stimulate potent biological responses!

Hmmmm! How to minimise risks and how to interpret outcomes?

No need too match the origin of ECM to the origin of cells. Podocytes are epithelial cells and physiologically bind basement membrane proteins such as laminin and collagen type IV.