# US 10,766,029 B2 · Device, platform, and assay for assessing cells

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- Number: US 10,766,029 B2
- Document ID: US10766029B2
- Status: Granted
- Kind: B2
- Application number: 15/551,007
- Filed: 2016-02-22
- Published: 2018-08-23
- Granted: 2020-09-08
- Earliest claimed: 2015-02-20
- Assignees: Cells for Cells S.A.
- Applicants: Cells for Cells S.A.
- Inventors: Juan Pablo Acevedo Cox
- Web page: https://patentgrep.com/patent/US10766029B2

## CPC

- B01L 3/5025: {for parallel transport of multiple samples}
- B01L 3/5027: {by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip}
- G01N 33/5008: {for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics}
- G01N 33/5029: {on cell motility}
- B01L 2200/0694: Creating chemical gradients in a fluid
- B01L 2300/069: Absorbents; Gels to retain a fluid
- B01L 2300/0816: Cards, e.g. flat sample carriers usually with flow in two horizontal directions
- B01L 2300/0864: comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
- B01L 2400/0409: centrifugal forces
- B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)

## Abstract

Devices are for assessing the migration response in the presence of a stable encapsulated gradient of a factor or factor combination, and quantifying the adherence response inside micro-channels in the presence of different factors. A platform is for obtaining information relating to migration score or the quantification of adhered cells through use of the devices, and it allows this information to be used to assess therapeutic potential. A method quantifies the cells migration response and the cell adherence response.

## Claims

### Claim 1

A device adapted to assess cell migration responses of a population of cells, the device comprising:
a) a cell load chamber;
b) at least two migration assay lanes or lines formed by two parallel hydrogel side bands and allowing migration, wherein the migration assay lanes or lines are parallel, wherein a migration area in each assay lane is delimited in a lower part by the two side bands and material capable of supporting the two side bands and allowing migration, wherein the assay lanes or lines are open in an upper part and the migration is produced on the material; and
c) wherein the hydrogel comprises an encapsulated migration inducing factor.

### Claim 2 (depends on 1)

The device according to claim 1, wherein the assay lines or lanes have a width of about 150 μm.

### Claim 3 (depends on 1)

The device according to claim 1, wherein the assay lanes or lines are spaced at a distance apart to form at least one assay channel.

### Claim 4 (depends on 1)

The device according to claim 1, wherein the assay channel has a width of about 50 μm, the assay channel has a height of about 50 μm and the assay channel has a length of about 10 mm.

### Claim 5 (depends on 1)

The device according to claim 1, wherein the encapsulated factor gradient comprises a concentration range of migration inducing factor from about 1 nM to 400 nM.

### Claim 6 (depends on 5)

The device according to claim 5, wherein the migration or adherence inducing factor is selected from the group consisting of Thrombin (F2), Interleukin 8 (IL-8, CXCL8), Factor derivative from 1α (SDF-1α, CXCL12) cell stroma, Wnt11, Wnt3, Leptin (LEP), Interleukin-13 (IL-13), Angiotensin II (ANGII), Melanoma cell adherence molecule (MCAM, CD146), Interleukin 2 (IL-2), Fibroblast growth factor 1 (FGF-1), Fibroblast growth factor 2 (FGF-2), Low molecular weight hyaluronic acid (LMWHA), Beta transforming growth factor (TGF-beta), Vascular endothelial growth factor (VEGF-B and VEGF-A), Lysophosphatidic acid, Regulated upon Activation, Normal T-cell Expressed, and Secreted (RANTES, CCL5), Interferon gamma-induced protein 10 (CXCL10, IP-10), Monocyte 1 chemoattractant protein 1 (MCP1, CCL2), Macrophage inflammatory protein 1α (MIP1α, CCL3), Macrophage inflammatory protein-1β (MIP-1β, CCL4), Chemokine (C—C motif) ligand 7 (CCL7), Macrophage inflammatory protein-3-beta (MIP-3-beta, CCL19), Chemokine (C—C motif) ligand 21 (CCL21), Chemokine (C—C motif) ligand 25CCL25, Lymphocyte B chemoattractant B (CXCL13), Chemokine (C—X—C motif) ligand 16 (CXCL16), Tumor necrosis factor-α (TNF-α), Hepatocytes growth factor (HGF), Epidermal growth factor (EGF), Platelet derivative growth factor (PDGF), Insulin growth factor (IGF), Angiopoietin-1 (ANGPT1), and Granulocyte colony stimulating factor (G-CSF), or combinations thereof.

### Claim 7 (depends on 1)

The device according to claim 1, wherein the encapsulating factor gradient is released upon addition of a release factor.

### Claim 8 (depends on 7)

The device according to claim 7, wherein the release factor is a protease.

### Claim 9 (depends on 8)

The device according to claim 8, wherein the protease is a collagenase.

### Claim 10 (depends on 1)

The device according to claim 1, wherein the device is adapted to assess cell migration response of a population of cells selected from the group consisting of mesenchymal stem cells, early mesenchymal/stromal precursor cells, adipose tissue-derived stem cells, Muse-AT cells, hematopoietic cells, hematopoietic stem cells, platelets, Kupffer cells, osteoclasts, megakaryocytes, granulocytes, NK cells, endothelial precursor or progenitor cells, pluripotent cells, CD34+ cells, Stro-1+ cells, Stro-3+ cells, CD29+ cells, CD166+ cells, Thy-1+ or CD90+ stem cells, CD44+ cells, immune cells, monocytes, leukocytes, lymphocytes, Band T cells, NK cells, macrophages, neutrophil leukocytes, neutrophils, neutrophil granulocytes, adult and embryo stem cells, endoderm mesenchymal stem cells (MSCs), mesoderm MSC, ectoderm MSC, early mesenchymal/stromal precursor, adipose tissue-derived stromal/stem cells, multipotent stem cells, adipocytes, osteocytes, chondrocytes, myoblasts, cardiomiocytes, astrocytes, and neuronal/glial cell lineages.

### Claim 11 (depends on 1)

The device according to claim 1, wherein the hydro-gel comprises gelatin, hyaluronic acid, alginate, agarose, chitose, gellan gum, collagen, collagen based hydrogel, high methacrylated salmon gelatin at 10%, polyethylene glycol, polyethylene acid, polyvynylpirrolidone, polyacrylamide, polymetyl methacrylate, polyethylene glycol dyacrilate based formula (PEGDA), pentaerythritol triacrylate (PETA), acrylic acid, acrylamide, or combinations thereof.

### Claim 12 (depends on 11)

The device according to claim 11, wherein the hydro-gel comprises collagen based hydrogel.

### Claim 13 (depends on 1)

The device according to claim 1, wherein the hydro-gel further comprises a photo initiator comprising 2-hydroxy-4′-(2-hydroxyethoxy)-2-methylpropiophenone or 2.2-dimethoxy-2-phenyl acetophenone.

### Claim 14 (depends on 1)

The device according to claim 1, which is adapted to assess cell adherence response of a population of cells, the device comprising:
a) a micro-channel circuit for perfusion of a liquid medium;
b) an internal substrate limiting channel edges;
c) at least one entry for a liquid medium injection; and
d) at least one entry for sample cells.

### Claim 15 (depends on 1)

A method to quantify cell migration response of a population of cells, the method comprising the steps of:
providing the cells to the device according to claim 1;
incubating the cells under conditions to facilitate migration;
calculating migration distance of cells contained in a sample to generate at least one score;
incorporating the at least one score in a statistical mathematical model specific to a cell type and pathology;
obtaining a correlation between the factor defined and therapeutic quality level; and
calculating a general performance value and therapeutic quality of the cell sample.

### Claim 16 (depends on 1)

A method of using the device according to claim 1, comprising assessing therapeutic characteristics of a sample stored in a cell bank.

### Claim 17 (depends on 1)

A method of using the device according to claim 1, comprising performing quality control with the device before therapy and after thawing a cell sample.

### Claim 18 (depends on 1)

A method of using the device according to claim 1, comprising assessing cell migration in presence of factors related to a rate of success in different treatments.

### Claim 19 (depends on 1)

A method of using the device according to claim 1, comprising calculating therapeutic variability of a cell sample or product according to cell's activity in the device.

### Claim 20 (depends on 1)

A method of using the device according to claim 1, comprising calculating cell doses to be administered to a patient during treatment.

### Claim 21 (depends on 1)

A method of using the device according to claim 1, comprising assessing cell migration in presence of factors related to a rate of success in kidney damage recovery.