Laser-based technique for controlled damage of mesenchymal cell spheroids: a first step in studying reparationin vitro

Abstract

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.

Influence of gonadotropins on ovarian follicle growth and developmentin vivoandin vitro

“…The selection of the optimal follicular culture conditions needed to obtain mature oocytes in vitro requires a complex approach. It is necessary to take into account the mechanical and chemical surroundings of the cells, metabolism and paracrine interactions (Filatov et al ., 2015; Kosheleva et al ., 2016). To obtain a greater number of mature oocytes is necessary to apply all the factors (including gonadotropin treatment) that are essential for normal follicle growth and development.…”

Cell spheroids as a versatile research platform: formation mechanisms, high throughput production, characterization and applications

“…In the case of spheroid production, effective transfer oxygen is observed for cell aggregates with thicknesses below 200 µm [131]. Therefore, the production of spheroids with small diameters, around 200 µm, is recommended for application in tissue engineering to ensure non-limiting mass transfer conditions [132].…”

Evaluation of the anti-oxidative and ROS scavenging properties of biomaterials coated with epigallocatechin gallate for tissue engineering

“…Necrotic core formation or external damage can lead to contraction and eruption of spheroids and thus structural deformation of a spheroid can serve as an indicator of the integrity of the spheroid. For example, extensive DNA damage was observed in multicellular tumor spheroids (MCTSs) cultured with H 2 O 2 (0.1-5 mM) for 1 hr, resulting in structural changes [51] , while laser irradiation of MSC spheroids caused destruction from the outside of the spheroid [52] . Our results indicate that the presence of E-PF protected against ROS damage.…”