Nonxenogeneic Growth and Retinal Differentiation of Human Induced Pluripotent Stem Cells

Abstract

Abstract Human induced pluripotent stem cells (hiPSCs) possess tremendous potential for the field of regenerative medicine because of their ability to differentiate into any cell type of the body. Such ability has profound implications for translational medicine, because these cells have been implicated for use in cell replacement, disease modeling, and pharmacological screening. However, the translation of established methods for deriving retinal cell types from hiPSCs has been hindered by the use of xenogeneic products for their growth and differentiation. Thus, the ability to derive retinal cell types in the absence of xenogeneic products would represent a significant advancement. The following studies were therefore undertaken to test the ability of hiPSCs to give rise to retinal cells under nonxenogeneic conditions. hiPSCs were maintained in traditional, feeder-free, or xeno-free culture conditions, and their ability to differentiate to a retinal fate was tested. Upon differentiation under all three conditions, cells acquired advancing features of retinal development, eventually yielding cell types of the mature retina. Reverse transcription-polymerase chain reaction and immunocytochemistry confirmed early trends in gene and protein expression patterns in xeno-free derived hiPSCs similar to those in cells derived in mouse embryonic fibroblasts and in feeder-free conditions. Results from this study demonstrate that hiPSCs can be maintained and directed to differentiate into retinal cell types under nonxenogeneic conditions, similar to cells derived using current xenogeneic methodologies. The demonstration of this capability will facilitate future efforts to develop hiPSC-based therapies for retinal disorders and also help to advance in vitro studies of human retinal development.