US 12,139,740 B2
Method for determining the invasive potential of a tumor cell
Manuel Thery, Paris (FR); Yoran Margaron, Voiron (FR); and Odile Filhol-Cochet, Claix (FR)
Assigned to COMMISSARIAT A L'ENERGIE ATOMIQUE ET AUX ENERGIES ALTERNATIVES, Paris (FR); and INSERM (INSTITUT NATIONAL DE LA SANTÉ ET DE LA RECHERCHE MÉDICALE), Paris (FR)
Appl. No. 16/615,418
Filed by COMMISSARIAT A L'ENERGIE ATOMIQUE ET AUX ENERGIES ALTERNATIVES, Paris (FR); and INSERM (INSTITUT NATIONAL DE LA SANTÉ ET DE LA RECHERCHE MÉDICALE), Paris (FR)
PCT Filed May 30, 2018, PCT No. PCT/EP2018/064129
§ 371(c)(1), (2) Date Nov. 21, 2019,
PCT Pub. No. WO2018/219980, PCT Pub. Date Dec. 6, 2018.
Claims priority of application No. 17305633 (EP), filed on May 31, 2017.
Prior Publication US 2020/0102590 A1, Apr. 2, 2020
Int. Cl. C12Q 1/02 (2006.01); C12M 1/34 (2006.01); C12N 5/09 (2010.01); G01N 33/50 (2006.01)
CPC C12Q 1/02 (2013.01) [C12M 41/46 (2013.01); C12N 5/0693 (2013.01); G01N 33/5011 (2013.01)] 14 Claims
 
1. A method for predicting the invasive potential of a tumor cell, comprising:
providing a culture device comprising a planar substrate and at least ten sets of two adhesive patterns for culturing adherent cells on said substrate, wherein the two adhesive patterns are sufficiently separated from each other by an essentially non-adhesive intercalating area for preventing a cell on a first adhesive pattern to reach another adhesive pattern; and, the area covered by the two adhesive patterns and the intercalating area is sufficient to adhere two animal cells, and wherein each set of two adhesive patterns is isolated by a cytophobic surrounding region;
seeding at least one tumor cell on said sets of two adhesive patterns;
culturing said tumor cell so as to obtain one cell doublet from one cell division on said sets of two adhesive patterns;
measuring at least three parameters on said sets of two adhesive patterns for said tumor cell, said at least three parameters comprising:
the internuclear distance for each set of two adhesive patterns, which is the length between the nucleus of two cells in the doublet on each set of two adhesive patterns;
the cell-cell connectivity for each set of two adhesive patterns; which is the difference between the area of the convex envelope surrounding the cell doublet and the area of the cell doublet;
and
the percentage of highly polarized cells to the total cell population on said sets of two adhesive patterns, wherein the highly polarized cells are cells having a centrosome located in close proximity of the cell-cell junction, in a triangular area defined as:
|y|≤(x−0.5)*tan 60
with 0.5≤x≤2
wherein x is the position of the centrosome on a nucleus-nucleus axis pointing toward the intercellular junction and y is the position of the centrosome on an axis perpendicular to the nucleus-nucleus axis, the unit being the radius of the nucleus;
the percentage of highly polarized cells is the ratio of the number of cells having a high polarization as defined above by the total number of cells*100; and
determining the tumor cell invasive potential based on the at least three measured parameters for the at least 10 cell doublets, by comparison with reference cells with a predetermined invasive potential, by performing statistical analysis.