The present invention relates, in general, to tissue engineering and, in particular, to a method of extending the lifespan of cells suitable for use in vascular engineering.
The present invention relates to a method of producing vascular grafts. The method comprises introducing into cells an expression vector that comprises a nucleic acid sequence encoding a lifespan extending and/or immortalizing gene, or functional portion or variant thereof, under conditions such that the sequence is expressed and the protein product of the gene, or functional portion or variant thereof, is produced. These cells can then be employed in the engineering of vascular grafts.
Cells suitable for use in the invention include SMCs, epithelial cells (of which endothelial cells (ECs) are a subset), fibroblasts, pericytes, cardiomyocytes, and nervous system cells. The cells can be fully differentiated vascular cells isolated from an individual. Alternatively, the cells can be derived from stem cells or progenitor cells that are cultured under conditions such that they differentiate into vascular cells. Advantageously, the cells are human cells derived from the graft recipient. Typically, the cells are non-neonatal SMCs derived from an individual of any age.
Cells can be harvested from a patient from a peripheral arterial or venous branch biopsy. The biopsy can be conducted anywhere on the patient's body that allows access to peripheral vasculature; the upper arm and lower leg are two examples. The surgical technique used can be that described for temporal artery biopsy (Albertini et al, Dermatalogic Surgery 25: 501-508 (1999) ). Doppler ultrasound or other means of imaging the vasculature (such as digital subtraction angiography) can be used to locate the peripheral vessel segment of interest. Under local anesthetic, the skin,
MCKEE J; NIKLASON LAURA; COUNTER CHRISTOPHER
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