Method for producing multi-layered microfluidized bed device for growing cell cultures in dynamic conditions

摘要

The notification shall cover the production of a multi-layered microfluid device for dynamic tissue production consisting of at least six layers, including: bottom and age, adhesive layer, at least one auxiliary glass layer, at least one elastomer functional layer, and at least one hydro-gel layer, which is characterised by the production in the first stage, on the basis of the pre-designed maps of each layer of the device, of its components -vascular and connecting channels and microchannels, revision, transition and positioning holes and breeding chambers,partitions and tanks- for this purpose, the elastomer layer is prepared by coating the carrier with an elastomer layer and solidifies, and then engraves and cuts the structure by irradiation with a concentrated laser beam; auxiliary glass layers, the adhesive layer and the bottom and age of the micro-device are engraved and cut into the structure by laser-focused irradiation of the material sheets; in the second stage, the auxiliary glass layers and the elastomer functional layers in the assembly are combined without the use of adhesive, each consisting of one glass layer and one elastomer layer on the vehicle constituting the temporary assembly position;in the third stage, the liquid hydrogel pre-prepared and reconstituted is introduced through the revision holes into the breeding chambers of the teams,and then disconnects the temporary base of the assembly, thereby engraving the vascular microchannels in the exposed layer, and in the partitions the connecting channels, by means of laser beam-focused illumination; in the fourth stage, one or more assemblies are connected to each other or to the bottom without the use of glue, and with age by means of adhesive layer; in the fifth stage, the micro-system connects with hoses that lead and drain liquids using glue; in the sixth stage, the micro-system is cleaned and washed; in the seventh stage, the suspension of cells of any type is introduced into the micro-system either once or repeatedly,at intervals enabling them to be immobilised on the surface of microchannels; in the eighth stage, cell culture shall be carried out under flow conditions in order to form a network of vessels inserted into artificial tissue.