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Temporary Microwells – simple and efficient way to homogeneous cell distribution on cell arrays

Abstract

The invention combines several advantages of conventional systems and offers a simple possibility to realize "temporary microwells" on a cell substrate. In addition, the new arrays ensure homogeneous distribution of transfection material and cells and are also suitable for solid phase transfection.

Background

Cell arrays are used in biological and medical research, e.g. to identify target structures for drugs, or in industrial drug development.

Problem

Cell arrays are used to carry out different studies on many cells simultaneously. In solid phase transfection, for exam­ple, transfection reagents are applied locally to a solid car­rier material and dried. The cells, which are subsequently distributed as homogeneously as possible on the entire carrier, are transfected at these spots. A homogeneous dis­tribution is decisive for comparative analysis.
In general, however, cells do not show the desired homo­geneous distribution after seeding, but often accumulate at the edge (‘coffee ring effect’) or in the center of the array (‘Marangoni effect’).

Solution

As part of a project funded by the Baden-Württemberg Stiftung gGmbH, researchers at the BioQuant research institute (University of Heidelberg) have developed a simple method, which allows ‘temporary microwells’ to be created on cell substrates. This method enables homo­geneous cell distribution with minimum technical effort. At the same time, it prevents sample crosstalk and cross-conta­mina­tion.
This innovative technology is also easy to implement: Prior to cell seeding, two grids of different pore sizes are placed on top of each other and integrated into the plate. Based on the combination of the grids, whose pore sizes are per­fectly positioned in relation to each other and to the spot sizes, temporary microstructures are formed on the large-format carrier material. These ‘temporary microwells’ spe­cifically restrict cell and reagent movement. After cell seed­ing and grid removal, parallel processing of the cells can be resumed without interruption.

Preferred form of cell arrangement based on the present invention: Solid carrier (bottom), a first net (center) on the carrier surface and a second, finer net (top) on the first net [BioQuant, University of Heidelberg].
Preferred form of cell arrangement based on the present invention: Solid carrier (bottom), a first net (center) on the carrier surface and a second, finer net (top) on the first net [BioQuant, University of Heidelberg].

Advantages

  • Effective avoidance of
    • inhomogeneous cell distribution on any substrates
    • sample crosstalk
    • cross-contamination on cell arrays
  • Parallel treatment using reagents is not inhibited by permanent microwells
  • Enhanced analysis
  • Development of new fields of application
  • Price optimization

Application

This invention makes it possible to produce cell arrays that combine the advantages of several conventional sys­tems (multi-well plates and microarrays). In addition, the new arrays ensure homogeneous distribution of trans­fected materials and cells. They can also be used for solid phase transfection.

Exposé
Contact
Anne Böse, M.Sc.
TLB GmbH
Ettlinger Straße 25
76137 Karlsruhe | Germany
Phone +49 721-79004-0
boese(at)tlb.de | www.tlb.de
Development Status
Prototype / TRL5
Patent Situation
EP (WO 2017/129367 A1) pending
Reference ID
15/022TLB
Service
TLB GmbH manages inventions until they are marketable and offers companies opportunities for license and collaboration agreements.