Biology and Medicine

Abstract

Isolation of a novel population of multipotent stem cells from epidermal layer of human skin

AB Balaji, Kaiser Jamil, G Maruthiram and CM Habibulla

Fore skin samples were collected after surgical excision from boys below 7 years of age from Deccan Medical College –Hyderabad, in cold saline (40 C) in sterile containers. These skin samples were cut into small pieces and incubated in trypsinised phosphate buffer saline (PBS) overnight to dissociate epidermis from the dermis. The epidermal layer which got detached was lifted and taken for preparing the cell suspension. Viability of cells was 88% with PI staining. About 50 μl of cell suspension was aliquoted into vials (105 cells/per vial ) and incubated with 2 μl of the antibodies. Various biomarkers of stem cells such as CD34 – one of the marker’s of skin stem cell, CD 49f & CD29 - common stem cell markers, CD45 Lymphocytic marker, CD90 (Thy-1) skin stem cell marker and CD105 endothelial marker and CD56 (NCAM) neural adherent marker were used . Using these cell surface markers of stem cells, cytometry was performed on a FACS Calibur with sorter (BD Biosciences). These immunophenotypic markers with Cellquest software expressed the stem cells characteristics. FACS analyses gave different profiles of stem cells /progenitors of mesenchymal, hematopoietic and neural progenitors. These expressed profiles indicated that its progenitor might have the multipotent and or pluripotent character. By using embryonic markers, we could select a population of stem cells to give multipotent or pluripotent cells that could have the capacity to regenerate skin cells for application in burn cases, wounds, deep burns and non-healing wounds. In conclusion, we present here for the first time the isolation of pluripotent / multipotent skin stem cells from human foreskin biopsy samples and its potential applications for various applications.