Hello all;
I am going to isolate, culture and do quality assays for Human Dermal Fibroblasts (HDFs).
I checked the literature review BUT STILL WOULD LIKE TO GET YOUR OPINIONS as this will be my very first time and some of you are experts, have done it before many times.
If you could help me with quality assays for HDFs ( such as immunophenotyping, some special protein staining...etc) I would appreciate it.
Thank you
You can check several parameters, for example: mitochondrial mass and membrane potential by Mitotracker and/or Mitosox, autophagy flux by a bafilomycin assay, senescence by sudan black staining, proliferation ratio and viability by MTT assay... There are plenty aproximations and techniques to test on HDFs. You should focus on what you want to see.
Thank you Juan Miguel Suárez-Rivero
well.. I am more into immunophenotyping. Therefore, I am looking for the right markers of HDFs. As far as I see they share markers with MSCs as they are CFU-F. But just want to see which markers the others use to understand If I chose the right ones.
Article Human Dermal Fibroblasts Demonstrate Positive Immunostaining...
Table 1. Primary antibodies used in this study.
Cell morphology, adherence rate, and cell viability testing
Flow cytometry analyses of the fibroblast-specific antigen CD90
Growth performance testing through multiple passages up to 10 or 15 population doublings (PD) under culture conditions without antibiotics and antimycotics (see table below).
Testing for the absence of HIV-1, HIV-2, HBV, HCV, HTLV-1, HTLV-2 and microbial contaminants (fungi, bacteria, and mycoplasma)
A detailed certificate of analysis (CoA) for each lot is available for download.
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