FSBSI Institute of General Pathology and Pathophysiology, 8 Baltiyskaya St, Moscow 125315, Russian Federation Faculty of Biology, Lomonosov Moscow State University, 12-1 Leninskie Gory, Moscow 119234, Russian Federation
Joint Institute for High Temperatures of the Russian Academy of Sciences, 13 Bld 2, Izhorskaya St., Moscow 125412, Russian Federation.
Biol Open. 2016 Jul 15;5(7):993-1000. doi: 10.1242/bio.017145.
Modern techniques of laser microsurgery of cell spheroids were used to develop a new simple reproducible model for studying repair and regeneration in vitro Nanosecond laser pulses (wavelength 355 nm, frequency 100 Hz, pulse duration 2 ns) were applied to perform a microdissection of the outer and the inner zones of human bone marrow multipotent mesenchymal stromal cells (BM MMSC) spheroids. To achieve effective dissection and preservation of spheroid viability, the energy of laser pulses was optimized and adjusted in the range 7-9 μJ. After microdissection, the edges of the wound surface opened and the angular opening reached a value of more than 180°. The destruction of the initial spheroid structure was observed in the wound area, with surviving cells changing their shape into a round one. Partial restoration of a spheroid form took place in the first six hours. The complete structure restoration accompanying the reparative processes occurred gradually over seven days due to remodelling of surviving cells.
采用现代激光微手术技术对细胞球体进行处理，开发出一种新的简单、可重现的体外修复和再生研究模型。纳秒激光脉冲（波长 355nm，频率 100Hz，脉冲持续时间 2ns）用于对外层和内层的人骨髓多能间充质基质细胞（BM MMSC）球体进行微切割。为了实现有效的切割和保持球体活力，激光脉冲的能量在 7-9μJ 的范围内进行了优化和调整。微切割后，伤口表面的边缘张开，角度开口达到 180°以上。在伤口区域观察到初始球体结构的破坏，存活的细胞形状变为圆形。在最初的六个小时内，球体形状部分得到恢复。由于存活细胞的重塑，在七天内逐渐发生完整结构的修复伴随修复过程。
