Encapsulation of trabecular meshwork mesenchymal stem cell using microfluidic system for differentiation into neuron-like cells.

Abstract

Objective: Neurodegenerative diseases are a class of nervous system disorders characterized by progressive neuronal degeneration and loss of function. Among emerging therapeutic approaches, microfluidic-enabled stem cell encapsulation and transplantation has gained recognition as a promising strategy for mitigating neuronal damage. In this work, human mesenchymal stem cells (MSCs), isolated from trabecular meshwork (TM) tissue, were successfully encapsulated and differentiated into neural-like cells via a microfluidic platform to demonstrate their potential for neural repair applications.

Materials and methods: The isolated mesenchymal stem cells were cultured on a microfluidic system (fabricated by soft lithography methods) and treated with medium containing DMEM supplemented with RA, IBMX, and forskolin for 7 days. Quantitative PCR (qPCR) were used to analyze differentiated TM-MSC and their expression of neural-like specific markers such as Nestin and b-tubulin 3.

Results: qPCR analysis revealed the presence of genes characteristic of neural cells (Nestin and β-tubulin 3) in cells differentiated both within a microfluidic system and on traditional tissue culture plates (TCPS). qPCR result showed that cells on 1.5% alginate showed higher expression of β-tubulin 3 compared to those on 1% alginate, 2% alginate, and TCPS (p < 0.0001). In contrast, Nestin expression showed no statistically significant differences across all pairwise comparisons (p > 0.05 for all groups).

Conclusion: The findings indicate that mesenchymal stem cells derived from the trabecular meshwork (TM-MSCs) may serve as promising candidates for cell-based therapeutic strategies. Furthermore, the microfluidic platform implemented in this study exhibits potential utility as a delivery vehicle for TM-MSCs in therapeutic interventions targeting neurological disorders.