Cellular Engineering Technologies (CET) has submitted this proposal in response to RFA-GM- 19-001. CET has proposed a direct Phase II SBIR application to create more reproducible human induced pluripotent stem cells (iPSCs) and create methods for growing, maintaining, and authenticating iPSCs. A major challenge in iPSC manufacturing and subsequent differentiation is the emergence of genetic instability that result from non-random chromosomal mutations. Genetic instability results in clonal expansion of genetic variants that increases iPSC heterogeneity. The experimental variables that promote genetic instability are not well understood. Yet, oncogene-dependent reprogramming and prolong cell culturing are clearly linked to genetic instability. Moreover, prior iPSC reprogramming methods adapted for preclinical research have not been optimized to mitigate against the infectious, inflammatory, neoplastic and genetic risks for cell therapy. Thus, iPSC reprogramming should be standardized to include non-integrating, virus-free and oncogene-free methods, which would offer reproducible iPSC in adherent and suspension cells. This milestone would mitigate oncogenic and viral effects that could reduce genetic instability in iPSC manufacturing and differentiation. Further, iPSC reproducibility and differentiation would improve if growth factors displayed fully human posttranslational modification (PTM). While bacterial-manufactured growth factors and non-human glycosylated peptides and proteins are ubiquitous in the stem cell field, they exhibit differential bioactivity than their native human counterparts. Thus, using growth factors that lack a fully human PTM may amplify the genetic instability and distort cell phenotype of iPSC and differentiated cells, particularly for multiple differentiation steps that require multiple growth factors. CET is a biotechnology company with a diverse pipeline of human somatic stem cells and a first-in-class non-integrating, feeder-free, virus-free and oncogene-free iPSC reprogramming approach that has been validated and published for adherent cells and suspension cells. Moreover, CET is the sole source manufacturer of select postnatal stem cells. These capabilities allowed CET to obtain immortalized human postnatal stem cells designed for biologic bioprocessing of fully human PTM. Thus, CET is poised to develop iPSC and differentiated cells through manufacturing processes that mitigate genetic instability. The focus of this proposal will be to develop a manufacturing platform to create GLP and GMP-grade iPSC with the least amount of genetic instability even after subsequent neuroprogenitor cell differentiation.