Skin Tissue Engineering & Morphology Core
The Skin Tissue Engineering & Morphology (STEM) Core is lead by director Kathleen J. Green, PhD, and co-directed by Bethany E. Perez-White, PhD. STEM Core facilitates fundamental research in skin biology by providing technological expertise and resources to support the culture of normal human and mouse epidermal keratinocytes, primary melanocyte and dermal fibroblast cultures, as well as co-cultured skin cell 3-D models. The core helps foster new collaborations among groups with the common goal of understanding the biology of skin cells.
Human Epidermal Keratinocytes
The core provides a reliable source of highly purified preparations of human epidermal keratinocytes isolated from individual or pooled, neonatal foreskin. Cells are supplied at a low passage number in either low calcium/serum-free culture medium, a fibroblast-feeder layer culture system or frozen ampoules. Training in keratinocyte isolation and keratinocyte media at bulk rates are also available to users.
Human Epidermal Keratinocytes from Patients with Skin Disease
On a case-by-case basis, the core isolates and amplifies human keratinocytes from biopsies of adults, including patients diagnosed with various skin diseases. The availability depends on consent to biopsy; utilization of the disease-specific keratinocytes for research requires IRB-approval. Cells are supplied at a low passage number on a fibroblast-feeder layer culture system or as frozen ampoules. Limited numbers of frozen ampoules generated from previously acquired patient material with certain diseases may be available on request.
Human Dermal Fibroblasts
Human dermal fibroblasts are isolated from individual or pooled, neonatal foreskin or patient skin biopsies. Cells are provided at a low passage number in either culture medium or frozen ampoules.
Human melanocytes are isolated from neonatal foreskin and provided at a low passage number in a commercial, serum-free culture medium system.
Murine Epidermal Keratinocytes
Keratinocytes isolated from the skin or tail of newborn and adult control and genetically modified mice are supplied as short- or long-term cultures in either low calcium/serum-free culture media or frozen ampoules.
Organotypic Raft Cultures
The core provides service, training and supplies necessary for generation of three-dimensional cultures of human epidermis. These so-called “raft” cultures are formed by plating primary control or disease keratinocytes on a collagen substrate infused with primary or immortalized fibroblasts, that are lifted to place the collagen “plug” onto an air-medium interface. Upon request, primary human melanocytes or melanoma cell lines can be co-cultured with epidermal keratinocytes in the 3-D raft model. Cultures take approximately nine to 12 days to mature into a stratified epidermal equivalent. Rat tail collagen, pre-cast collagen lattices and other supplies necessary for raft culture generation will also be supplied.
Immortalization of Human Epidermal Keratinocytes
Primary keratinocytes are immortalized using the HPV elements of the herpes simplex virus, HPV-16, and can be supplied in either culture media or frozen ampoules. In addition, keratinocyte lifespan can be extended by pharmacological inhibition of Rho-ROCK signaling for cultures maintained on a fibroblast feeder layer system.
Kathleen J. Green, PhD
Joseph L. Mayberry, Sr., Professor of Pathology and Toxicology
Professor of Pathology and Dermatology
Bethany E. Perez-White, PhD
Associate Core Director
Assistant Professor of Dermatology
For more information, please reach out to the people below.
Bethany Perez White
Associate Core Director
Senior Research Technologist