Flow cytometric sorting and analysis of human epidermal stem cell candidates

Abstract

ESC (epidermal stem cells) play a central role in the regeneration of human epidermis. These cells are also responsible for wound healing and neoplasm formation. Efficient isolation of ESC allows their use in medicine and pharmacy as well as in basic science. Cultured keratinocytes and ESC may be used as biological dressing in burn injuries, chronic wounds and hereditary disorders. Therefore, the isolation and characterization of ESC have been goals in biomedical science. Here, we present a flow cytometric method for the isolation and analysis of human ESC candidates. The strategy presented for the isolation of ESC combines previously proposed enzymatic digestion and FACS-sorting of the obtained cell suspension that utilizes morphological features, integrin-beta1 expression and Rh123 (Rhodamine 123) accumulation of the cells. We also performed a flow cytometric analysis of sorted cells using a cell tracer.

Transcriptional profile of in vitro expanded human epidermal progenitor cells for the treatment of non-healing wounds

“…Grafting the autologous cells is highly suitable since there is no risk of graft rejection and viral transmission. However, this method has many limitations related to the time required for cell expansion (usually 2-4 weeks), differentiation process in vitro (decreased level of progenitor cells) and the lack of specific environmental cell niche [24,25]. The presented study involved elaborating a novel therapy protocol enabling effective and successful clinical application as well as molecular and phenotypic analysis of in vitro cultured cells including transcriptomic profiling of in vitro cultured epidermal progenitor cells.…”

“…The signal is mediated by unoccupied beta-1 integrin that could also lead to apoptotic death in a lack of specific environmental milieu [33]. Thus, it is crucial to routinely monitor the level of beta-1 integrin in cultured cells and the alternative integrin subunits of heterodimers [25].…”

α6 Integrin (α6high)/Transferrin Receptor (CD71)low Keratinocyte Stem Cells Are More Potent for Generating Reconstructed Skin Epidermis Than Rapid Adherent Cells

“…For example, integrin α6-antibody coupled to fluorochrome or to magnetic beads allows the isolation of positive cells by flow cytometry [21] or under a magnetic field, respectively. Combination of β1-integrin and Rhodamine 123A [22], as well as β1-integrin and Desmoglein 3 [10], or β1-integrin and K1/K10 [23], would allow isolation of epidermal stem cells candidate. A cell population α6 high /CD71 low sorted by flow cytometry [8] was also proposed to isolate KSC.…”

Current Status of Healthy Human Skin Models: Can Histone Deacetylase Inhibitors Potentially Improve the Present Replacement Models?

“…For detailed information on the development of skin epithelium, regulation of keratinocyte differentiation and skin morphology in general we refer to existing articles and reviews [3,55,56,57,58,59,60,61,62,63,64,65,66,67,68,69,70,71]. …”