Arrays of discrete cell culture microenvironments, method of making such arrays and uses thereof
a cell culture microenvironment and array technology, applied in the field of arrays of discrete cell culture microenvironments, can solve the problems of inability to faithfully reproduce certain key physiological features of in vivo cell culture, inability to independently control the key matrix parameters, and inability to accurately reproduce in vivo gene expression patterns and cellular phenotypes, etc., to achieve the effect of changing the degradability
Inactive Publication Date: 2016-04-28
ECOLE POLYTECHNIQUE FEDERALE DE LAUSANNE (EPFL)
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Benefits of technology
The patent is about a method for creating matrixes that can change their properties over time when exposed to proteases, such as those produced by cells. This allows for the efficient growth and migration of cells in a 3D environment. The method involves adding peptides to the matrix that are sensitive to proteases and adjusting the content of polymers and other components to achieve the desired degradation characteristics. The resulting matrix can also contain biologically active molecules that can influence cell behavior. Additionally, the patent discusses the challenge of controlling the stiffness of gels used for 3D cell culture. The inventors have developed a solution that allows for independent control of stiffness and degradability, making it easier to create optimal microenvironments for cell culture.
Problems solved by technology
Two-dimensional cell culture systems have the drawback of not allowing for gene expression patterns and cellular phenotypes that closely resemble those found in vivo.
Further, they cannot be expected to faithfully reproduce certain key physiological features of the in vivo cell culture microenvironment, notably spatial constraints, proteolytic remodeling, stiffness-mediated mechanotransduction and appropriate mode of presentation of ligands.
Naturally derived 3D cell culture systems have poorly defined compositions and show batch to batch variation, which makes it impossible to alter their properties in systematic ways and to independently control their key matrix parameters.
Method used
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[0086]Hereinbelow, embodiments of the invention will be presented in greater detail, by means of embodiments.
[0087]Generation of Arrays of 3D Signaling Cell Culture Microenvironments from a Tool-Kit of Molecular Building Blocks
[0088]To realize the three-dimensional screening of cellular microenvironments according to the present invention it has proven useful to engineer a biomaterials system composed of a library of molecular building blocks which can be independently mixed and then cross-linked in the presence of cells to potentially form a huge diversity of three-dimensional cell microenvironments with distinct and independently controllable properties. It was found that the invention can preferably be carried out on the basis of synthetic hydrogels as biomimetic three-dimensional cell microenvironments with very well defined biochemical and mechanical properties (Lutolf and Hubbell, 2005). Preferably, the coagulation enzyme activated transglutaminase factor XIIIa (FXIIIa) is use...
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Abstract
The invention pertains to a combinatorial method of identifying the hydrogel formulations controlling phenotype and fate of difficult-to-culture cell types.
Description
[0001]The present invention pertains to arrays of discrete volumes of cell culture microenvironments possessing different properties, i.e. different mechanical and / or biochemical properties (i.e. cell adhesion, signalling, degradability) influencing cell phenotype and fate, i.e. proliferation / self-renewal, colony formation, differentiation and / or migration, methods of making such arrays, a kit-of-parts for making such arrays and a combinatorial method of testing the influence of hydrogel formulations on cell growth behaviour, in particular of difficult-to-culture cell types such as stem cells.BACKGROUND OF THE INVENTION[0002]Two-dimensional cell culture systems as well as naturally derived three-dimensional cell culture systems as models to elucidate complex cell behaviour, notably in the field of cancer or stem cell research, have been known for many years.[0003]Two-dimensional cell culture systems have the drawback of not allowing for gene expression patterns and cellular phenotyp...
Claims
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IPC IPC(8): G01N33/50
CPCG01N33/5073G01N33/502G01N33/5005C12N5/00C12N5/06
InventorLUTOLF, MATTHIASRANGA, ADRIAN
OwnerECOLE POLYTECHNIQUE FEDERALE DE LAUSANNE (EPFL)