Head and neck stem-like cells were identified by cell sorting for the surface markers CD44-phycoerythrin (PE) (BD Biosciences, San Jose, CA, USA) [33 (link),34 (link)], CD117-fluorescein isothiocyanate (FITC) (BD Biosciences, San Jose, CA, USA), CD133-allophycocyanin (APC) (Miltenyi Biotec, Bergisch Gladbach, Germany) [2 (link),35 (link),36 (link)], and the intracellular marker ALDH (aldehyde dehydrogenase) [37 (link),38 (link),39 (link)]. ALDH activity was detected using an Aldefluor Kit (StemCell Technologies, Durham, NC, USA) according to the manufacturer’s instructions. Flow cytometry analysis was performed on FACSAria II (BD Biosciences, Mountain View, CA, USA) and FACSMelody (BD Biosciences, Mountain View, CA, USA) equipment. Table 2 summarizes all the markers used for immunophenotyping and cell sorting from primary cultures and cell lines.
Cd44 phycoerythrin pe
Manufactured by BD
Sourced in United States
CD44-phycoerythrin (PE) is a fluorescently-labeled antibody that binds to the CD44 cell surface receptor. CD44 is a glycoprotein involved in cell-cell interactions, cell adhesion, and migration. The phycoerythrin (PE) fluorescent dye allows for the detection and quantification of CD44-expressing cells using flow cytometry or other fluorescence-based techniques.
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Lab products found in correlation
Cd133 allophycocyanin apc, by Miltenyi Biotec (1 mentions)
Facsmelody, by BD (1 mentions)
Aldefluor kit, by STEMCELL (1 mentions)
Facsaria 2, by BD (1 mentions)
Cd34 pe, by BD (1 mentions)
Alizarin red, by Yuanye Bio-Technology (1 mentions)
Oil red o, by Yuanye Bio-Technology (1 mentions)
Cd105 fluorescein isothiocyanate fitc, by BD (1 mentions)
Cd45 fitc, by BD (1 mentions)
2 protocols using cd44 phycoerythrin pe
1
Head and Neck Stem-Like Cell Identification
2
Characterization and Differentiation of Stem Cells
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The cells (1x105/cm2) suspensions were incubated with antibodies against CD44-phycoerythrin (PE), CD34-PE, CD105-fluorescein isothiocyanate (FITC), and CD45-FITC, respectively (all from BD Bio-Sciences, San Jose, CA, USA), in the dark for 30 min at 4 °C. The fluorescence intensity was measured using flow cytometry. To evaluate the differentiation potential, the stem cells were inoculated into 12-well plates at a density of 5×103/cm2 according to the manufacturer’s protocol for the differentiation-inducing medium. Osteogenesis, adipogenesis, and chondrogenesis media (differentiation kits from Gibco, Thermo Fisher Scientific, Waltham, MA, USA), were used to culture the cells for 21 days (osteogenesis) and 28 days (adipogenesis, chondrogenesis). The osteogenesis, adipogenesis, and chondrogenic differentiation abilities of stem cells loaded with or without SPION@PDA NPs were detected by Alizarin Red, Oil Red O, and Alcian blue staining kits, respectively (Shanghai Yuanye Bio-Technology Co., Ltd, Shanghai, China).
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