did u try coating the substrate with ECM proteins to facilitate attachment? and did u plasma treat the substrates to make it hydrophilic :) these could help :)

Hi, 

Maybe you already aware that there is two level of Petri dishes. One is cheaper, normally we use it for agar gel, bacteria culture and etc; the other is tissue culture level, which is normally treated with polystyrene. Polystyrene generates a hydrophobic condition, which is good for the cell attachment. 

I remember when I culture HUVEC, or want to culture the cell on the cover glass, I will perform gelatin coating on the surface of the substrate. I think the gelatin solution (for cell culture level) can be available at a reasonable price.  

* of course, perform under sterile condition. 

1. Allow gelatin solution to completely liquefy at 37°C.

2. Coat culture surface with 5-10 µL gelatin solution/cm2 (i.e., 0.1-0.2 mg/cm2 gelatin).

* normally, I will reuse the gelatin.

3. Allow to dry at least 2 hours before introducing cells and medium.

Cell culture grade plastics are chemically treated. The treated culture surface are positively charged. Proteins on cell membrane are mostly negative charged. That is why cells are easily attached to cell culture grade plastics. Whereas hydrogel film is much complicated and glass surface are more neutral.

Thank you all for your valuable feedback.

@Darren wong, Can you please share more details about how to coat with ECM and treatment with plasma?

@Shulin low, you are probably right about the polystyrene, however, taking this into account, do you think all hydrophobic surfaces are provide better attachment to cells? I cant proceed with the gelatin coating as i have to proceed in to in-vivo studies. 

@james shen,Do you think coating the films with positively charged amino acids will facilitate cellular attachment?

Hi Fayyaz, if the film is already positively charged, it is difficult that the amino acid will bind to, but you can try.

Hi, 

I am not sure why you can't proceed to gelatin coating even you need to proceed to in-vivo studies. I don't know any other method, sorry about that. Good luck

@James, thank you very much for your answer. I will consider these suggestions in the future experiments after discussing with my supervisor. 

@Shulin Low, The reason i cant proceed with the gelatin or any other coating of animal origin is because of the sensitive nature of experiment. The in-vivo studies are about the transplant immunology where you dont want anything of zoonotic potential. That is the reason why collagen, fibronectin and gelatin coatings are not being tested. I'm grateful for your suggestions. I will definitely reply to this post after experimenting different things and let you all know what worked for me. 

Probably you have solved your problem, but this page helps you to understand the adhesive substrates.

Evolution of Cell Culture Surfaces

By: John A. Ryan, BioFiles 2008, 3.8, 21.

https://www.sigmaaldrich.com/china-mainland/zh/technical-documents/articles/biofiles/evolution-of-cell.html