Developing a Method for the Phenotypic Preservation of Nucleus Pulposus Cells for Autologous Cell Implantation - ANAT 396 Undergraduate Research Project Application Form

Supervisor's Name: Lisbet Haglund

Supervisor's Email: lisbet.haglund [at] mcgill.ca

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Supervisor's Website:

Supervisor's department: Experimental surgery

Course number: ANAT 396 (Anatomy and Cell Biology)

Term: Winter 2013-2014

Project start date: Monday, January 6, 2014

Project end date: Friday, April 11, 2014

Project title: Developing a Method for the Phenotypic Preservation of Nucleus Pulposus Cells for Autologous Cell Implantation

Project description (50-100 words suggested): The process of autologous cells implantation, which is currently investigated for its potential therapeutic effects on Disk Degenerative Disease (DDD), involves the culture of primary cells from a patients Nucleus Pulposus (NP) tissue. The growth surfaces currently used for these cultures are silicon covered plastic plates treated with collagen type I. However, it has been observed that NP cells tend to dedifferentiate and revert to a more fibrotic phenotype when grown in these types of static culture environments. This is characterised by (but not limited to) a decrease in SOX-9, collagen type II, aggrecan, Chondroadherin (CHAD) and Glucoseaminoglycan (GAG) production and an increase in collagen type I production. This is a hindrance to the efficiency of the procedure, since these cell products are critical for the biomechanical functionality of the intervertebral disc. We hypothesized that this was due to a disruption of the NP cells’ environment during the passaging process from one plate to another. We attempted to address this issue by cultivating primary NP cells (from bovine samples) on an expanding culture medium, which will reduce the need to disrupt cell colonies and therefore preserve their phenotypic characteristics. RNA will be extracted from the freshly isolated bovine NP cells and at different time intervals during culture of NP cells on silicon coated plates and on an expanding culture medium. These samples will then be assessed for expression of the aforementioned phenotypic criteria. Media and RNA samples will be extracted for analysis through DMMB (for net GAG content) and Q-PCR (for gene expression of collagen type I and II, aggrecan, CHAD and SOX-9). RNA isolated from the freshly isolated NP cells will provide a baseline level of gene expression, to which samples from both culture methods will be compared to assess phenotypic preservation.

Prerequisite: 1 term completed at McGill + CGPA of 3.0 or higher; or permission of instructor.

Grading scheme (The final report must be worth at least 50% of final grade): The results will be presented in a final paper which will count for 50% of the final course mark. The remaining 50% will be based on participation as assessed by the supervisor.

Project status: This project is taken. The professor has no more '396' projects this term.

Ethics, safety, and training: Supervisors are responsible for the ethics and safety compliance of undergraduate students. This project involves: biohazardous substances, handling chemicals.