Process Development for Scalable Manufacturing of hESC-Derived Cardiomyocytes

associated with cancer treatments, or by parathyroidectomy, commonly lead to hypoparathyroidism. Patients with hypoparathyroidism suffer from serious complications including painful tetany and muscle spasms, chronic kidney problems such as nephrocalcinosis or even renal failure, arrhythmias and signifi cant CNS problems. Therefore, restoration of parathyroid gland function would greatly improve the quality of life of an individual suffering from hypoparathyroidism. The reimplantation of parathyroid glands following thyroid surgery has been somewhat successful for restoring parathyroid function; however, this is not an approach that can always be used and must often be supplemented pharmacologically. The recent generation of hiPS cells has opened the door to the possibility of developing autologous cellular therapies with patient derived cells that could be benefi cial for individuals with hypoparathyroidism. Reprogramming patient-specifi c human somatic cells into iPS cells with pluripotence features similar to those of embryonic stem (ES) cells creates an opportunity for autologous repair and/or regeneration of tissues/ organs that have been damaged by disease or therapeutic interventions with minimal prospect for immune rejection. To achieve this goal, an in vitro culture protocol was developed to generate functional parathyroid gland epithelial cell progenitors from patient specifi c hiPS cells. The protocol was designed based on human embryological developmental stages to enhance the potential for generating a mature functional gland epithelium that will actively secret parathyroid hormone in response to environmental stimuli. This study assesses the directed differentiation of hiPS cells into defi nitive endoderm, anterior foregut endoderm (AFE), and parathyroid progenitor cells. The experiments demonstrate that brief treatment of the cells with high doses of Activin A in the early stages of iPS cell differentiation promotes endodermal differentiation, and that the readdition of high doses of Activin A to a parathyroid differentiation cocktail that includes Sonic HedgeHog (SHH) after the detection of pharyngeal pouches, leads to the increased expression of parathyroid markers. This work has been supported by Pennsylvania Cystic Fibrosis, Inc and the Hearing Research Institute.