Laser Research Centre, Faculty of Health Science, University of Johannesburg.
Laser Research Centre, Faculty of Health Science, University of Johannesburg;
J Vis Exp. 2024 Apr 5(206). doi: 10.3791/66616.
Adipose-derived stem cells (ADSCs), possessing multipotent mesenchymal characteristics akin to stem cells, are frequently employed in regenerative medicine due to their capacity for a diverse range of cell differentiation and their ability to enhance migration, proliferation, and mitigate inflammation. However, ADSCs often face challenges in survival and engraftment within wounds, primarily due to unfavorable inflammatory conditions. To address this issue, hydrogels have been developed to sustain ADSC viability in wounds and expedite the wound healing process. Here, we aimed to assess the synergistic impact of photobiomodulation (PBM) on ADSC proliferation and cytotoxicity within a 3D cell culture framework. Immortalized ADSCs were seeded into 10 µL hydrogels at a density of 2.5 x 10 cells and subjected to irradiation using 525 nm and 825 nm diodes at fluencies of 5 J/cm and 10 J/cm. Morphological changes, cytotoxicity, and proliferation were evaluated at 24 h and 10 days post-PBM exposure. The ADSCs exhibited a rounded morphology and were dispersed throughout the gel as individual cells or spheroid aggregates. Importantly, both PBM and 3D culture framework displayed no cytotoxic effects on the cells, while PBM significantly enhanced the proliferation rates of ADSCs. In conclusion, this study demonstrates the use of hydrogel as a suitable 3D environment for ADSC culture and introduces PBM as a significant augmentation strategy, particularly addressing the slow proliferation rates associated with 3D cell culture.
脂肪来源干细胞（ADSCs）具有多能间充质特性，类似于干细胞，由于其能够进行多种细胞分化，并且能够增强迁移、增殖和减轻炎症的能力，因此经常被用于再生医学。然而，ADSCs 在伤口内的生存和植入常常面临挑战，主要是由于炎症条件不利。为了解决这个问题，已经开发出水凝胶来维持伤口内 ADSC 的活力并加速伤口愈合过程。在这里，我们旨在评估光生物调节（PBM）对 3D 细胞培养框架内 ADSC 增殖和细胞毒性的协同影响。将永生化 ADSC 以 2.5x10 个细胞的密度接种到 10µL 水凝胶中，并使用 525nm 和 825nm 二极管以 5 J/cm 和 10 J/cm 的通量进行辐照。在 PBM 暴露后 24 小时和 10 天评估形态变化、细胞毒性和增殖。ADSCs 呈圆形形态，作为单个细胞或球体聚集物分散在凝胶中。重要的是，PBM 和 3D 培养框架都对细胞没有细胞毒性作用，而 PBM 显著提高了 ADSC 的增殖率。总之，本研究表明水凝胶可作为 ADSC 培养的合适 3D 环境，并引入 PBM 作为一种重要的增强策略，特别是针对 3D 细胞培养中与增殖缓慢相关的问题。
