TY - DATA T1 - Aldefluor protocol to sort keratinocytes stem cells from skin PY - 2017/12/20 AU - Samuel Marcos Ribeiro Noronha AU - Alfredo Gragnani AU - Thiago Antônio Calado Pereira AU - Silvana Aparecida Alves Correa AU - Jessica Bonucci AU - Lydia Masako Ferreira UR - https://scielo.figshare.com/articles/dataset/Aldefluor_protocol_to_sort_keratinocytes_stem_cells_from_skin/5718877 DO - 10.6084/m9.figshare.5718877.v1 L4 - https://ndownloader.figshare.com/files/10042009 L4 - https://ndownloader.figshare.com/files/10042018 L4 - https://ndownloader.figshare.com/files/10042021 L4 - https://ndownloader.figshare.com/files/10042033 L4 - https://ndownloader.figshare.com/files/10042039 L4 - https://ndownloader.figshare.com/files/10042063 KW - Keratinocytes KW - Flow Cytometry KW - Adult Stem Cells. N2 - Abstract Purpose: To investigate the use Aldefluor® and N, N - Dimethylaminobenzaldehyde (DEAB) to design a protocol to sort keratinocyte stem cells from cultured keratinocytes from burned patients. Methods: Activated Aldefluor® aliquots were prepared and maintained at temperature between 2 to 8°C, or stored at -20°C. Next, the cells were collected following the standard protocol of sample preparation. Results: Best results were obtained with Aldefluor® 1.5µl and DEAB 15 µl for 1 x 106 cells, incubated at 37°C for 15 minutes. Flow cytometer range for keratinocyte stem cells separation was evaluated. There were 14.8% of stem cells separated in one sample of keratinocyte culture used to pattern the protocol. After being defined the ideal concentration, the same test pattern was performed in other keratinocyte samples. We observed a final mean of 10.8%. Conclusion: Aldefluor® has been shown as a favorable marking of epidermal keratinocyte stem cells for subsequent separation on a flow cytometer, with detection of 10.8% of epidermal keratinocyte stem cells, in this protocol. ER -