Dear all,
I have a problem in culture of HDFs. Every time I seed them, they detach from the surface of the flask, or even they don't adhere at all. Consequently, precipitation occur without any change in color of the medium, even after a week. I tested everything and used other companies for medium (DMEM), serum (FBS), and the flask (poly L_Lysine coated), but the same result achieved. the PH of the medium was 7.9. It's virtually high but not as much as to bring about detachment of the cells. I've cultured any other cell lines but I'd never experienced this problem. What's your advice?
Dear Maryam Sardari ,
Look at the following data:
https://pubmed.ncbi.nlm.nih.gov/23996453/
Article Cytogentic analysis of human dermal fibroblasts (HDFs) in ea...
https://www.atcc.org/products/pcs-201-012
What is human dermal fibroblasts?
Dermal fibroblasts are the main cell type present in skin connective tissue (dermis). Fibroblasts interact with epidermal cells during hair development and in intrafollicular skin. Moreover, they play an essential role during cutaneous wound healing and in bioengineering of skin.
Isolation and Culture of Human Dermal Fibroblasts - PubMed
https://pubmed.ncbi.nlm.nih.gov › ...
Search for: What is human dermal fibroblasts?
What do dermal fibroblasts do?
Dermal fibroblasts produce and organize the extracellular matrix of the dermis. They also communicate with each other and other cell types. Fibroblasts play a crucial role in regulating skin physiology and cutaneous wound repair.
Dermal Fibroblast - an overview | ScienceDirect Topics
https://www.sciencedirect.com › topics › engineering › de...
Search for: What do dermal fibroblasts do?
Where do human dermal fibroblasts come from?
Dermal fibroblasts are derived from mesenchymal stem cells within the body. Like corneal fibroblasts, dermal fibroblast proliferation can be stimulated by the presence of fibroblast growth factor (FGF).
Dermal fibroblast - Wikipedia
https://en.wikipedia.org/wiki/Dermal_fibroblast
Search for: Where do human dermal fibroblasts come from?
How do humans get fibroblasts?
Fibroblasts in patients will maintain all genetic background during reprogramming into induced pluripotent stem cells. In spite of their large use, fibroblasts are obtained after an invasive procedure, a superficial punch skin biopsy, collected under patient's local anesthesia.
An extensive discussion related to dermal fibroblasts:
https://www.researchgate.net/post/Problem-with-human-fibroblast-culture
Typically, when fibroblasts do not attach, it is either something wrong with the cells (for example, cells are dead) or something inhibiting attachment (e.g., residual trypsin or something inhibiting attachment). It is not clear what is going on in your case. One thing you could try is to use collagen-coated flasks, which fibroblasts directly attach rather than ECM proteins in the serum.
I have worked with HDFs but never faced this problem. You can try differently coated flasks or coat them yourself with more fibroblast-friendly coating. Also test for mycoplasmas.
- Badges
- Science topic
- Similar topics
- Computer Science and Engineering
- Computer Graphics and Animation
- 3D