Isolating stem cells from skin: designing a novel highly efficient non-enzymatic approach

Abstract

Stem cells are undifferentiated elements capable to acquire a specific cellular phenotype under the influence of specific stimuli, thus being involved in tissue integrity and maintenance. In the skin tissue self-renewal and wound healing after injury is a complex process, especially in adulthood, due to the aging process and the continuous exposure to damaging agents. The importance of stem cells in regenerative medicine is well known and defining or improving their isolation methods is therefore a primary and crucial step. In the present paper we present a novel method to isolate stem cells from human skin, including the involvement of a novel medium for the maintenance and expansion of in vitro cultures. The biopsies were mechanically digested and put in culture. The migrating cells were positive selected with magnetic cell sorting, characterized by flow-cytometry analysis, and viability detected by MTT assay. Cells exhibited a mesenchymal phenotype, as demonstrated by the positive acquirement of an osteogenic or adipogenic phenotype when cultured in specific conditioned media. Taken together our results disclose a novel method for culturing and expanding stem cells from skin and pave the way for future clinical applications in tissue regeneration.

Smart Nanofibers with Natural Extracts Prevent Senescence Patterning in a Dynamic Cell Culture Model of Human Skin

“…In the skin, the fundamental resource for self-renewal is represented by skin stem cells, which, as well as stem cells derived from other tissues, are able to differentiate towards a selected cell phenotype, under specific stimuli [31,[35][36][37]. Indeed, skin stem cells are closely related to the activity of the other cell types resident in the tissue.…”

“…Human skin stem cells were obtained from biopsies of adult male and female patients after ethics committee approval (Ethical Clearance N. 0021565/2018, 22/03/2018-Commissione Etica CNR). They were isolated and cultured as previously described [31]. Human skin fibroblast 1 (HFF1) and Human skin keratinocytes (HACAT) were purchased from ATCC (Manassas, VA, USA) and cultured in a Dulbecco's modified Eagle's Medium (DMEM) low-glucose medium (Life Technologies, Carlsbad, CA, USA), supplemented with 10% fetal bovine serum (FBS Life Technologies), 2 mM l-glutamine (Euroclone, Milano, Italy) and 1% of penicillin/streptomycin (Euroclone) [32].…”

Natural Compounds and PCL Nanofibers: A Novel Tool to Counteract Stem Cell Senescence

“…Human SSCs and Human skin fibroblasts 1 (HFF1), obtained as previously described [12,55], were cultured in a DMEM low-glucose medium (Life Technologies, Carlsbad, CA, USA), supplemented with 10% fetal bovine serum (FBS Life Technologies, Carlsbad, CA, USA), 2 mM/L-glutamine (Euroclone, Milano, Italy), and 1% penicillin/streptomycin (Euroclone, Milano, Italy).…”

“…These properties can be significantly reduced after UV exposure and oxidative-stressrelated dysfunctions [5]. Stem cells are able to undergo self-renewal or commit toward a specific differentiated phenotype upon physical or chemical stimuli [6][7][8][9][10], exhibiting different plasticity and differentiation potential, according to their tissue source [6,[10][11][12][13]. Skin comprises several stem cell types, showing different roles, properties, and differentiation capabilities [14,15].…”

“…Skin comprises several stem cell types, showing different roles, properties, and differentiation capabilities [14,15]. In particular, skin stem cells (SSCs), a mesenchymal cell population, are able to undergo different fates due to their asymmetric division: one cell can maintain the undifferentiated condition in the niche, while the other can be recruited into becoming a differentiated cell [12]. SSCs were found both in the epidermis and dermis.…”