Hi Handan,

we performed human skin fibroblast isolation mincing the tissue in very small pieces, let them attach on plastiv for 24-48 hours with a very small medium volume (just covering a 35mm plate) and then we fill the dish in order to cover the culture with D-MEM 10% FBS. After 5-7 days you should see cells coming out from the skin pieces cultured.

  Good luck

Dear Handan,

Usually we use little pieces of skin, we let them attach in 35mm petri dish for 24-48 hrs (make sure the dermis is in direct contact with the culture surface). We use DMEM with 20% serum and change media every 2 days. Just pay attention not to move the culture dish too much, in order to avoid detachment of the tissue. In less than 2 weeks you should see fibroblasts coming out.

Best,

Ximena 

Hello Handan,

I need some more information to answer to your question:

Are your fibroblasts from Human or Mouse biopsies ? 

What are the size of the biopsies ?

Why do you use 30% of FBS? It seems huge for me , and can you tell me the CO2 % you use in your incubators ?

Best

Yannick

Thank you for all answers.

I used human fibroblast biopsy and cut it into 2 mm small pieces. But I guess, my fault was not to contact dermis and culture disk surface. The other things that I applied were the same as Cesare's suggestions (except for 30% of FBS). I used 30% of FBS on some suggestions in ResearchGate. Last I used 0,5 % CO2.

Best regards