publication . Article . 2017

Aldefluor protocol to sort keratinocytes stem cells from skin

Noronha, Samuel Marcos Ribeiro; Gragnani, Alfredo; Pereira, Thiago Antônio Calado; Correa, Silvana Aparecida Alves; Bonucci, Jessica; Ferreira, Lydia Masako;
Open Access
  • Published: 01 Nov 2017 Journal: Acta Cirurgica Brasileira, volume 32, pages 984-994 (issn: 0102-8650, eissn: 1678-2674, Copyright policy)
  • Publisher: FapUNIFESP (SciELO)
Abstract
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 kerat...
Subjects
free text keywords: Keratinocytes, Flow Cytometry, Adult Stem Cells., Crystallography, Medicine, business.industry, business, Surgery, medicine.medical_specialty
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