Directed Differentiation of Dopamine-Secreting Cells from Nurr1/GPX1 Expressing Murine Embryonic Stem Cells Cultured on Matrigel-Coated PCL Scaffolds

Abstract

Parkinson's disease (PD) is a progressive neurological disorder characterized by a large number of motor and non-motor features and is known as the second most common neurodegenerative disorder after Alzheimer's disease. The hallmark pathology of PD is the damage and death of dopamine-producing neurons in the substantia-nigra of midbrain. Intrastriatal transplants of fetal mesencephalon derived DAergic neurons have provided proof-of-principle for the cell replacement strategy and have demonstrated reinnervation of the denervated striatum. However, ethical, technical, and practical limitations of deploying fetal DAergic neurons as the source for cell therapy in PD have ceased the spread of this procedure into clinical practice. Embryonic stem (ES) cells have emerged as a therapeutic alternative that can proliferate extensively and generate dopamine-producing neurons. To this extent and to surmount the obstacles related to embryonic neural cells, many investigations have focused on using pluripotent stem cells for the derivation of DAergic neurons. In the present study, a mouse embryonic stem (mES) R1 cell line was generated which could stably co-express Nurr1 (an essential transcription factor in DAergic neuron development) and GPX-1 (a neuroprotective enzyme against oxidative stress). The Nurr1/GPX-1-expressing ES cells (Nurr1/GPX-1-ES) were differentiated into DAergic-like cells via a three-dimensional culture environment consisting of Poly-ε-Caprolactone (PCL) nanofibrous scaffolds embedded by Matrigel (Mtg) in the presence of specific signaling molecules. DAergic neuron-specific genes were highly expressed in ES-derived DAergic neurons cultured and differentiated on PCL/Mtg scaffolds. Reverse-phase HPLC confirmed that the Nurr1/GPX-1-ES-cells differentiated on PCL/Mtg electrospun scaffolds could efficiently and exclusively secrete dopamine in response to stimulus. In conclusion, our results demonstrated that PCL/Matrigel nanofibrous scaffolds could efficiently support and promote the generation of functional DAergic-like cells from Nurr1/GPX-1-ES cells. The results of this study may have an impact on future tissue engineering for cell therapy of PD.

Three-dimensional hyaluronic acid hydrogel-based models for in vitro human iPSC-derived NPC culture and differentiation

“…18-20 Although Matrigel has been demonstrated as an effective CNS-related cell carrier, it cannot be employed for clinical usage due to its unknown growth factor composition and murine sarcoma origins. 21-23 Therefore, there is an unmet need to develop other component-defined hydrogel systems to support hiPSC-NPCs and to improve the controllability. A number of attempts have been made to develop modified hyaluronic acid, agarose, alginate, or peptides-based hydrogel materials for hiPSC encapsulation.…”

Materials for Neural Differentiation, Trans‐Differentiation, and Modeling of Neurological Disease

“…The results demonstrated that the Fmoc‐SAPs hydrogels are biocompatible in vivo and are utilized in neural grafts of intact brains . Terraf et al generated the mouse embryonic stem (mES) R1 cell line for the co‐expression of nuclear receptor‐related 1 protein (Nurr1) and GPX‐1. Poly‐ε‐caprolactone (PCL)/Matrigel nanofibrous scaffolds were used to establish the 3D environment .…”

“…Terraf et al generated the mouse embryonic stem (mES) R1 cell line for the co‐expression of nuclear receptor‐related 1 protein (Nurr1) and GPX‐1. Poly‐ε‐caprolactone (PCL)/Matrigel nanofibrous scaffolds were used to establish the 3D environment . The biocompatible nanofibrous scaffolds were fabricated by weaving the PCL solution with aluminum foil and were coated with gold .…”

“…Poly‐ε‐caprolactone (PCL)/Matrigel nanofibrous scaffolds were used to establish the 3D environment . The biocompatible nanofibrous scaffolds were fabricated by weaving the PCL solution with aluminum foil and were coated with gold . The results showed that the 3D culture environment could enhance the efficiency for differentiating oxidant‐resistant DAergic‐like cells, which were viewed as the source for PD cell treatment .…”

“…The biocompatible nanofibrous scaffolds were fabricated by weaving the PCL solution with aluminum foil and were coated with gold . The results showed that the 3D culture environment could enhance the efficiency for differentiating oxidant‐resistant DAergic‐like cells, which were viewed as the source for PD cell treatment . Electrospun biodegradable polymers were also selected as the scaffolds for promoting the generation of neural progenitor cells from the mouse ES cells .…”