Abstract 4929: Injection of Human Neural Stem Cell Line in a Mouse Hind Limb Ischemia Model Promotes Muscle Neovascularisation

Abstract

Objective: Recently, the human conditionally immortalized neural stem cell (hNSC) line, CTX0E03, has received approval from the UK MHRA for treatment, by direct intracerebral implantation, of stable stroke disability in elderly patients. We report here for the first time that intramuscular injection of the CTX0E03 hNSC line into a mouse model of hind-limb ischemia results in a repeatable significant, dose-dependent therapeutic improvement in blood flow. Methods: Unilateral limb ischemia was induced in immunodeficient (FoxnCD1 nu/nu ) mice. Suspensions of CTX0E03 cells were administered by injection into the ischemic limb adductor muscle. Five different doses of hNSCs were tested in two experiments (3 million; 1 million; 300,000 in Exp 1 and 300,000, 30,000 and 3,000 in Exp 2), with vehicle injections used as a control in both experiments (n=10/group). Laser Doppler measurements of the limbs were taken at Days 3, 7, 14 and 21. Capillary and arteriole density and absolute blood flow measurements (0.02 μ m fluorescent microspheres) were done on Day 21. Results: The three highest doses of CTX0E03 cells markedly increased the recovery of blood flow to the ischemic limb as measured by Doppler (Figure A ), adductor muscle blood flow (Figure B ) and capillary and arteriole density. The two lower doses, 30,000 and 3,000 cells produced no significant improvements. Histological analysis demonstrated CTX0E03 cell survival at Day 21 in some, but not all, of the implanted muscles. Conclusion: CTX0E03 cell therapy significantly improved clinically relevant aspects of hind limb ischemia in nude mice dose dependently by enhancing neovascularization.