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US 10,288,603 B2Granted
3D cell viability assay
AssigneeGreiner Bio-One North America, Inc.
InventorGlauco R. Souza
Life of this patent
Estimating the term…
Abstract
Cells are grown in 3D culture and topological features obtained by photomicrography are correlated to cell viability and cell cell interactions.
Claims16
A wound closure assay, comprising:
- a) perforating a plurality of samples of magnetized cell clusters levitating in a magnetic field or magnetically patterning a plurality of samples of magnetized cells levitating in a magnetic field to form levitating 3D cell clusters having a hole therein, one or more of said plurality of samples being control samples without a test agent, and one or more of said plurality of samples being test samples with one or more concentrations of said test agent;
- b) taking photomicrographs of said plurality of samples of said levitating 3D cell clusters with said hole at one or more times;
- and
- c) analyzing said photomicrographs to measure wound closure, wherein:
- wound
-
-
- closure
-
-
- (
- %
- )
- =
- 100
-
- %
- -
- (
- area
-
-
- of
-
-
- wound
- t
- area
-
-
- of
-
-
- wound
- t
-
-
- 0
- ×
- 100
-
- %
- )
- ,
- wherein said area of woundt0 is an area of said hole before said test agent is added and said area of woundt is an area of said hole at a given time t after addition of said test agent.
42 more elements
A cell assay comprising:
- a) culturing a plurality of samples of magnetized cells levitating in a magnetic field to form levitating 3D cell clusters, one or more of said plurality of samples being control samples without a test agent, and one or more of said plurality of samples being test samples with one or more concentrations of said test agent;
- b) taking photomicrographs of said plurality of samples of said levitating 3D cell clusters at one or more times;
- and
- c) analyzing said photomicrographs to measure fractal dimension (Df) and one or more of i) number of cell clusters, ii) cell cluster size, iii) total area of cell clusters, and iv) rate of cell cluster contraction;
- wherein said fractal dimension and said cell cluster size are directly proportional to cell viability and cell-cell interactions, and said number of cell clusters and said total area of cell clusters are inversely proportional to said cell viability and said cell-cell interactions.
2 more elements
A cell assay comprising:
- a) culturing a plurality of samples of levitating 3D cell cultures, one or more of said plurality of samples being control samples without a test agent, and one or more of said plurality of samples being test samples with one or more concentrations of said test agent;
- b) taking photomicrographs of said plurality of samples of said levitating 3D cell cultures at one or more times;
- c) analyzing said photomicrographs to measure fractal dimension (Df ) and one or more of:
- c-i) number of cell clusters,
- c-ii) cell cluster size,
- c-iii) total area of cell clusters, and
- c-iv) rate of cell cluster contraction;
- d) wherein said fractal dimension and said cell cluster size are directly proportional to cell viability and cell-cell interactions, and said number of cell clusters and said total area of cell clusters are inversely proportional to said cell viability and said cell-cell interactions;
- e) wherein said levitating 3D cell cultures are prepared by levitating cells with a composition comprising:
- e-i) negatively-charged nanoparticles;
- e-ii) positively-charged nanoparticles;
- and
- e-iii) support molecules,
- wherein one of e-i) and e-ii) includes magnetically responsive iron or iron oxide, and wherein said support molecules hold said negatively-charged nanoparticles and said positively-charged nanoparticles in an intimate admixture.
11 more elements
Dependencies are inferred from the claim text.