Investigation of characteristics of feather follicle stem cells and their regeneration potential.

Feather follicles have the extraordinary ability to regenerate and undergo molting cycles. Being tissue-specific stem cells, feather follicle stem cells (FFSCs) have a strong capacity for proliferation and are presumed to be progenitor cells for various epidermal organs. In order to characterize FFSCs and to understand how the feather epidermis and FFSCs produce such a reliable differentiation program resulting in the formation of complex feathers, We developed a culture scheme to select and expand FFSCs from chick feather follicles. FFSCs were examined with cell profiles, mutilpotential differentiation and immunocytochemical staining. FFSCs from a single clone were capable of self-renewal and proliferation. These cells expressed integrin ß1, CD49c, cytokeratin 15 (K15), cytokeratin 19 (K19) and a neural-genic cell marker, nestin, but not a teminal differentiation-related keratinocyte marker, cytokeratin 10 (K10). FFSCs could trans-differentiate into adipocytes, neurocytes and keratinocytes. The formation of micro-feather like structures ex-vivo also revealed the potential of regeneration. These results demonstrate that FFSCs possess the properties of stem/progenitor cells and may therefore serve as a useful model for studying mechanisms of stem cell differentiation and their involvement in organ regeneration.

Matrix interference in serum total thyroxin (T4) time-resolved fluorescence immunoassay (TRFIA) and its elimination with the use of streptavidin-biotin separation technique.

In our development of total serum thyroxin TRFIA using an immobilized second-antibody (S-Ab) as the separation agent, we observed a significant measurement bias caused by a matrix interference when the immobilized S-Ab had a relatively low binding capacity for the primary anti-T4 monoclonal antibody (McAb). Therefore, we employed a new separation system based on the highly active surface streptavidin and biotinylated anti-T4 McAb. Our results indicate that the matrix interference was removed and the assay performance was improved with the use of streptavidin-biotin separation technique. In our method, microwells were first coated with biotinylated BSA and then a streptavidin solution in the presence of 1% BSA was added to allow streptavidin to be immobilized via the pre-coated BSA-biotin. Surface streptavidin prepared in this protocol expressed a significantly increased binding capacity for the biotinylated anti-T4 McAb, compared to the passively adsorbed S-Ab for binding the original anti-T4 McAb. The immunoreactions between the biotinylated anti-T4 McAb and the T4 in the standard or sample or the europium-labeled T4-BSA conjugate mainly occurred in liquid solution, and then the immune complex was specifically trapped by the surface streptavidin and isolated from the free trace by washing. Serum TT4 TRFIA based on surface streptavidin was accurate, precise and economic, maintained all the merits of the directly immobilized surface antibodies.