Aldefluor protocol to sort keratinocytes stem cells from skin

Aldefluor protocol to sort keratinocytes stem cells from skin

Author Ribeiro Noronha, Samuel Marcos Autor UNIFESP Google Scholar
Gragnani, Alfredo Autor UNIFESP Google Scholar
Calado Pereira, Thiago Antonio Autor UNIFESP Google Scholar
Alves Correa, Silvana Aparecida Autor UNIFESP Google Scholar
Bonucci, Jessica Autor UNIFESP Google Scholar
Ferreira, Lydia Masako Autor UNIFESP Google Scholar
Abstract Purpose: To investigate the use Aldefluor r and N, N-Dimethylaminobenzaldehyde (DEAB) to design a protocol to sort keratinocyte stem cells from cultured keratinocytes from burned patients. Methods: Activated Aldefluor r aliquots were prepared and maintained at temperature between 2 to 8 degrees C, or stored at -20 degrees C. Next, the cells were collected following the standard protocol of sample preparation. Results: Best results were obtained with Aldefluor r 1.5 mu l and DEAB 15 mu l for 1 x 10(6) cells, incubated at 37 degrees 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 r 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.
Keywords Keratinocytes
Flow Cytometry
Adult Stem Cells
xmlui.dri2xhtml.METS-1.0.item-coverage Sao Paulo
Language English
Sponsor FAPESP
Grant number FAPESP: 2013/10905-0
Date 2017
Published in Acta Cirurgica Brasileira. Sao Paulo, v. 32, n. 11, p. 984-994, 2017.
ISSN 0102-8650 (Sherpa/Romeo, impact factor)
Publisher Acta Cirurgica Brasileira
Extent 984-994
Access rights Open access Open Access
Type Article
Web of Science ID WOS:000417846700010
SciELO ID S0102-86502017001100984 (statistics in SciELO)

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