Capturing avian somatic cells using feather pulp fibroblast culture as a non-invasive approach to biobanking...

Avian cells biobanking is an essential tool to safeguard the biodiversity of rare or endangered species. Reproductive germ cells are especially useful for species conservation as they can either be used directly as functional gametes or differentiated into functional gametes in vitro. However, obtaining and preserving avian germ cells is difficult and usually the number of breeding individuals and available surplus eggs are limited. In contrast, avian fibroblast cells are readily obtained using minimally invasive biopsies which makes them ideal for the establishment of genetic cellular resource biobanks for species in need. Improvements in fibroblast culture conditions will enable more extensive studies in physiology and genetics, among others, which are not currently available for most bird species, in addition to providing material for induced pluripotent stem cell generation. Here, we tested different culture protocols to improve the recovery and proliferation rates of fibroblasts derived from feather follicles of Red-and-green Macaw (Ara chloropterus, Psittacidae). We found that feathers collected during their initial growth phase (< 10 days after plucking) have the best potential for culturing and biobanking. We also determined that the inclusion of FGF-2 and IGF-1 in the culture media was decisive for their proliferation and viability. Under these conditions we were able to integrate the reprogramming factors Nanog and Pou5f3, resulting the generation of several fibroblast cell lines immortalized. These findings validated the use of growing feathers as an effective and non-invasive approach to obtain genetic material for avian biobanks with a wide range of applications.