Course Description

In this half-semester laboratory module, students will: 

• Use advanced cell culture techniques to explore key topics in tissue engineering 
• Create complex skin equivalent tissue, including mesenchymal/extracellular matrix and epithelial layers; evaluate experimental biomaterials for potential to improve process 
• Investigate angiogenic capacity of engineered tissue using in vivo system 
• Fix, embed, section and stain engineered tissue constructs for structural analysis 

Lecture Topics

• Tissue engineering: state-of-the art, future directions 
• Biology of Tissues: epithelial/mesenchymal interactions 
• Skin as Model Organ: epidermal/dermal structure and differentiation 
• Angiogenesis: significance for tissue engineering 
• Theory and practice of: cell culture and co-culture; angiogenic assays; histology: fixation, paraffin embedding, sectioning and staining 

Lab Topics

Week 1

Review cell culture techniques, lab safety, record keeping; Cast dermal layer of skin with cellular and acellular collagen components, plus experimental biomaterials; Initiate keratinocyte cultures.

Week 2

Seed keratinocytes on dermal layer; manipulate medium and culture conditions to induce differentiation.

Week 3

Use Matrigel to perform endothelial cell tube angiogenesis assays; Design experiments to assess angiogenic potential of unknown agents.

Weeks 4–7

Skin equivalent tissues matures. Test for ability of the tissue to support angiogenesis using a chick chorioallantoic membrane assay; Fix, paraffin embed, section and stain mature skin equivalent tissue for analysis of tissue differentiation and effects of biomaterials.

Prerequisites

The normal pre-requisite, BIT 466/566, is not offered Fall 2023. Students with prior animal cell culture experience, for example BIT 410/510, other coursework, or laboratory research experience may enroll with permission of instructor. Please contact Douglas Tremblay at dptrembl@ncsu.edu.