Year 3 + NCE

CIRM Progress Report Part A: Scientific Progress

I. Project Overview

During the past three years (36 months) we have successfully completed the milestones defined in the NGA for this grant. In brief, we have selected 1 clinically compliant ESC line H14 (and a back-up line H9), which have shown reproducible, efficient differentiation to dopaminergic neurons at lab scale. We have performed in vitro and in vivo characterization as defined in the NGA and guided by our discussion with our program officer at CIRM. We have determined the time point at which dopaminergic precursors (14 days after the NSC stage) can be frozen, shipped, thawed and transplanted without compromising their ability to mature and provide therapeutic benefit in animal models. Importantly, we have evaluated efficacy of cryopreserved dopaminergic precursors manufactured by the GMP-compatible process in a rodent PD model and shown functional recovery up to 6 months post transplantation as well as survival of dopaminergic neurons.

In the meanwhile we have successfully transferred the process of generating transplant ready dopaminergic neurons to the manufacture facilities at City of Hope (COH). They have adapted and optimized our protocols and have established GMP-compatible protocols for the culture of ESC-NSC and for differentiating NSC to Stage 3, Day 14 DA precursors for transplantation. During this reporting period (36 month), we have tested the equivalency of these lots and confirmed that lots manufactured at COH are consistent and similar to cells produced in the laboratory.

Our effort resulted in two important manuscripts in Cytotherapy:

1. Liu, Q., Pedersen, OZ., Peng, J., Couture, LA., Rao, MS., and Zeng, X. Optimizing dopaminergic differentiation of pluripotent stem cells for the manufacture of dopaminergic neurons for transplantation. Cytotherapy. 2013 Aug;15(8):999-1010.
2. Peng, J., Liu, Q., Rao, MS., and Zeng, X. Survival and engraftment of dopaminergic neurons manufactured by a GMP-compatible process. Cytotherapy. 2014 Sep;16(9):1305-12.