Scalable, safe and GMP-compatible production of embryonic stem cell derived retinal pigment epithelial cells

Regenerative medicine is an exponentially growing field that aims to regenerate a lost function, cell type or tissue due to damage, ageing or disease. Currently, more than 30,000 gene- and cell-based therapies have been or are being tested in clinical trials. Since the eye benefits from accessibility and a supposed to be immune privilege, many groups are exploring different strategies to treat diseases affecting this organ. Age-related macular degeneration (AMD), the leading cause of blindness in people aged over 65 years old, could be one of the first diseases treated with human pluripotent stem cells (hPSC)- derived therapies. This thesis has been focused on the development of a scalable, robust, defined and xeno-free protocol to differentiate hPSC into RPE-like cells, ensuring the safety of the obtained product through genomic, tumorigenicity and biodistribution studies. Finally, the differentiation of an in-house derived GMP-grade hESC line using a completely GMPcompliant protocol, together with the validation of a set of in-process and Quality Control tests has allowed to engage in conversations with the regulatory authorities to bring these cells closer to near clinical trials, and ultimately to AMD patients

Preclinical safety studies of human embryonic stem cell‐derived retinal pigment epithelial cells for the treatment of age‐related macular degeneration

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Publisher Correction: Identification of cell surface markers and establishment of monolayer differentiation to retinal pigment epithelial cells

An amendment to this paper has been published and can be accessed via a link at the top of the paper

Induction of functional dopamine neurons from human astrocytes in vitro and mouse astrocytes in a Parkinson's disease model

Cell replacement therapies for neurodegenerative disease have focused on transplantation of the cell types affected by the pathological process. Here we describe an alternative strategy for Parkinson's disease in which dopamine neurons are generated by direct conversion of astrocytes. Using three transcription factors, NEUROD1, ASCL1 and LMX1A, and the microRNA miR218, collectively designated NeAL218, we reprogram human astrocytes in vitro, and mouse astrocytes in vivo, into induced dopamine neurons (iDANs). Reprogramming efficiency in vitro is improved by small molecules that promote chromatin remodeling and activate the TGFβ, Shh and Wnt signaling pathways. The reprogramming efficiency of human astrocytes reaches up to 16%, resulting in iDANs with appropriate midbrain markers and excitability. In a mouse model of Parkinson's disease, NeAL218 alone reprograms adult striatal astrocytes into iDANs that are excitable and correct some aspects of motor behavior in vivo, including gait impairments. With further optimization, this approach may enable clinical therapies for Parkinson's disease by delivery of genes rather than cells