Neural priming of adipose-derived stem cells by cell-imprinted substrates*

Abstract

Cell-imprinting technology is a novel method for directing stem cell fate using substrates molded from target cells. Here, we fabricated and studied cell-imprinted substrates for neural priming in human adipose-derived stem cells in the absence of chemical cues. We molded polydimethylsiloxane (PDMS) silicone substrates on fixed differentiated neural progenitor cells (ReNcellTM VM). The ReNcellTM cell line consists of immortalized human neural progenitor cells that are capable to differentiate into neural cells. The fabricated cell-imprinted silicone substrates represent the geometrical micro-and nanotopology of the target cell morphology. During the molding procedure, no transfer of cellular proteins was detectable. In the first test with undifferentiated ReNcellTM VM cells, the cell-imprinted substrates could accelerate neural differentiation. With adipose-derived stem cells cultivated on the imprinted substrates, we observed modifications of cell morphology, shifting from spread to elongated shape. Both immunofluorescence and quantitative gene expression analysis showed upregulation of neural stem cell and early neuronal markers. Our study, for the first time, demonstrated the effectiveness of cell-imprinted substrates for neural priming of adipose-derived stem cells for regenerative medicine applications.

Computational and experimental studies of a cell-imprinted-based integrated microfluidic device for biomedical applications

“…Cell-imprinting technology is an innovative technique for directing stem cell fate by molding substrates from target cells. This method's functionality has been proven in previous research for various cells for different applications such as drug analysis and stem cell differentiation 2,[19][20][21][22][23]25,27,80 . The template cells are cultured and fixed on the cell culture plate's surface in this method.…”

“…A cell-imprinted substrate was fabricated from the chondrocyte shape as a physical stimulus for inducing chondrogenic differentiation in stem cells. Cardiomyogenic, tenogenic, osteogenic, kertainogenic, and Schwann cell differentiation in stem cells were also obtained with the imprinting method [19][20][21][22][23][24] . Also, the effect of physical topography on the cancer cells' response to the conventional anti-cancer drugs was investigated in 25 .…”

Molecular imprinting as a simple way for the long-term maintenance of the stemness and proliferation potential of adipose-derived stem cells: an in vitro study

“…Then the mold was peeled off from the tissue culture plate and rinsed thoroughly with 1 M NaOH solution for 30 minutes to eliminate any residual cell debris and existing chemicals from the imprinted substrates. 2,49,54,55,59 2.4. Surface characterization of PDMS substrates Scanning Electron Microscopy (SEM, Philips XL30, Netherlands) images of the substrates were evaluated based on previously published protocols.…”

Recent Developments in Surface Topography-Modulated Neurogenesis

“…6d) [56]. More recently, Ghazali et al choose a similar topography technique on adiposederived mesenchymal stem cells (ADSC) [57]. After a differentiation period of 14 days, ReNcell™ VM cells were fixed with 4% glutaraldehyde.…”

“…Based on those studies, cultured cells maintain a sensitivity to their environment that allows them to detect and respond to different topographies. Remarkably, the synergic effect caused by the combination of topography with other biophysical cues such as electrical Tissue topography Silica Silica BioReplication Cell morphology, focal adhesion and neuron-like cell differentiation [56] Cell topography PDMS Cell imprinting Cell morphology and early neuronal markers [57] stimulation, stiffness and biochemical molecules, was significantly more investigated. Yet, the underlying processes by which neurons, neuron-like cells and stem cells are capable of transducing such cues are not completely understood.…”