MIPT Cell Signaling Regulation Laboratory scientists have developed a new, low-cost and reproducible system for cell co-cultivation. This system is based on the polymerized BSA membrane. Its size and topography are determined by a template created with a 3D printer. The possibility of co-cultivation is realized using magnetic nanoparticles (NPs). The NPs are bound to the membrane, allowing them to remain afloat in a culture fluid using a constant magnetic field. The study was published in the journal Bioprinting.
To study how different cells affect each other, one must first separate them in space. The simplest cell co-cultivation model is based on the application of the so-called conditioned medium. Two groups of cells are grown separately, and then the culture medium that was used to grow one population is collected and used in another cell group. However, the problem with this model is that the short-lived molecules are not stable in a conditioned medium and do not have time to migrate to the recipient cell population. Thus, this results in the absence of feedback signals present under normal conditions
Clifford Grobstein achieved one of the most important achievements in the development of combined culture systems in 1953. He used permeable inserts with microporous membranes. Such systems for modeling changes in cellular phenotype are presented by the “Transwell System”. The main disadvantages of commercially available systems are the high price and the inability to independently reproduce them in the laboratory. However, this problem can be solved using 3D printing. This technology allows the production of custom-designed devices quickly and correctly for laboratory experiments with sufficient detail and accuracy.
MIPT Cell Signaling Regulation Laboratory researchers have developed their own low-cost, repeatable co-cultivation system.
Ilya Zubarev, Lead Investigator and Senior Research Fellow at the MIPT Cell Signaling Regulation Laboratory commented that they “decided to create a matrix based on a synaptic protein. Such systems are often used to simulate biological arrays. To realize the possibility of co-culturing different cells we chose an original solution that was not used Before, to keep the protein membrane above a certain height inside the vessel, we decided to saturate it with magnetic nanoparticles, and as the main component of the membrane, bovine serum albumin (BSA) was selected.This protein is non-toxic, widely available, and actively used in various fields of biology and, as a rule, is already available in any laboratory. ”
The possibility of co-cultivation is realized through the use of a magnetic support system of the floating membrane. For this purpose, magnetic nanoparticles are added to the membrane, and a system based on permanent magnets is placed on top of the dish. Cells cultured on the membrane retain their viability and can divide, the membrane can be fixed for histochemical or immunohistochemical staining, and cells can be separated from the membrane for further study. The cost of the finished membrane is about 1 US dollar, which is several times lower than commercially available analogues. The 3D printing method allows the manufacturing process to be modified flexibly and quickly according to the needs of a particular laboratory. These protein membranes can be manufactured in any laboratory using a 3D printer and widely available generic laboratory reagents.
Zubarev added that, “At the bottom of the Petri dish, as in the membrane, different types of cells grew. The membrane containing the cells was kept afloat in the culture medium. For several days, the cells exchanged signals with each other, after which it was from It is possible to assess their mutual effect. This co-cultivation system can be manufactured in any laboratory, and is a cost-effective alternative to commercial tabs of cell co-culture.”
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Minin et al., Development of a cell co-culturing system based on magnetic protein membranes, using an MSLA 3D printer, Bioprinting (2021). DOI: 10.1016 / j.bprint.2021.e00150
Presented by the Moscow Institute of Physics and Technology
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