Scalability and Safety Studies in Clinical-Grade Pluripotent-Derived Myogenic Progenitors for Therapeutic Application in DMD
Abstract
As proposed in the application, during the first year we performed a series of purification studies using CD54 antibody to identify the conditions for the optimal purification of iPAX7 human iPS cell-derived myogenic progenitors. These experiments resulted in the identification of the optimal ratio of antibody and microbeads for the MACS-mediated purification of our cell product with minimal residual of CD54-negative cells (Fig. 1-B). In addition, in order to identify the minimal media components required for the maintenance of their muscle regenerative potential, we tested multiple media conditions for the generation and expansion of CD54+ cells from iPAX7 iPS cells (Fig. 1C). These studies are ongoing as they require intramuscular transplantation in recipient animals followed by analysis 2 months post-injection. We observed that removal of horse serum has no impact on the proliferation and differentiation of CD54+ cells into muscle. To assess safety of human iPAX7 iPS cells, we performed karyotype analysis (Fig. 1D) and all genome sequencing using the 10X Genomics Chromium platform. Data are in the analysis pipeline and will include comparison to the genomic variants reported for the original cell line generated at NIH.
Document Details
- Document Type
- Technical Report
- Publication Date
- Oct 01, 2018
- Accession Number
- AD1074292
Entities
People
- Rita Perlingeiro
Organizations
- Regents of the University of Minnesota