# US 2015/0307848 A1 · Isolation and characterization of tumor cells using shear stress measurements

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- Number: US 2015/0307848 A1
- Document ID: US20150307848A1
- Status: Published application
- Kind: A1
- Application number: 14/713,241
- Filed: 2015-05-15
- Published: 2015-10-29
- Earliest claimed: 2011-03-24
- Assignees: University of Iowa Research Foundation
- Applicants: University of Iowa Research Foundation
- Inventors: Michael D. Henry; J. Matthew Barnes
- Web page: https://patentgrep.com/patent/US20150307848A1

## CPC

- G01N 1/40: Concentrating samples
- B01L 3/502715: {characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces}
- B01L 3/50273: {characterised by the means or forces applied to move the fluids}
- C12N 5/0693: {Tumour cells; Cancer cells}
- G01N 33/487: of liquid biological material
- G01N 33/5026: {on cell morphology}
- G01N 33/5044: {involving specific cell types}
- G01N 33/5091: {for testing the pathological state of an organism}
- G01N 33/575: for cancer
- B01L 2200/027: for microfluidic devices
- B01L 2200/0647: Handling flowable solids, e.g. microscopic beads, cells, particles
- B01L 2400/0475: specific mechanical means and fluid pressure
- C12N 2521/00: Culture process characterised by the use of hydrostatic pressure, flow or shear forces
- 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)
- B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

Methods for isolating viable cancer cells from a sample that comprises a mixture of cancerous cells and normal (non-cancerous) cells are provided. In the methods, a fluid preparation comprising a mixture of cancerous and normal cells is repeatedly exposed to fluid shear stresses, whereby the repeated exposure to the fluid shear stresses preferentially imparts fluid shear stress-resistance to the cancerous cells.

## Claims

### Claim at position 1

-18. (canceled)

### Claim 19

A method for purifying a fluid sample comprising extracellular calcium and cells of a mammalian subject, the cells comprising viable normal cells, the method comprising:
applying an initial pulse of fluid shear stress to the sample in vitro, wherein the initial pulse of fluid shear stress is sufficient to induce a fluid shear stress resistance in viable cancerous cells present in the sample such that the magnitude of shear stress required to kill the viable cancerous cells is greater than the magnitude of shear stress required to kill the viable normal cells; and
subsequently applying one or more additional pulses of fluid shear stress to the sample in vitro, said fluid shear stress sufficient to selectively kill the viable normal cells relative to the viable cancerous cells such that the ratio of the viable cancerous cells to the viable normal cells is increased.

### Claim 20 (depends on 19)

The method of claim 19, wherein the cells of the mammalian subject comprise lung cells.

### Claim 21 (depends on 20)

The method of claim 20, wherein the mammalian subject is a human.

### Claim 22 (depends on 19)

The method of claim 19, wherein the fluid sample is an in vitro fluid preparation.

### Claim 23 (depends on 22)

The method of claim 22, wherein the in vitro fluid preparation is prepared from a solid tissue biopsy.

### Claim 24 (depends on 19)

The method of claim 19, wherein the cells of the mammalian subject comprise the viable cancerous cells in addition to the viable normal cells.

### Claim 25 (depends on 19)

The method of claim 19, wherein the viable cancerous cells are viable cancerous epithelial cells.

### Claim 26 (depends on 19)

The method of claim 19, wherein the ratio of the viable cancerous cells to the viable normal cells in the sample is at least doubled.

### Claim 27 (depends on 19)

The method of claim 19, wherein the percent viability for the viable cancerous cells is at least 40% and the percent viability for the viable normal cells is no greater than 20% substantially immediately following the application of the initial and additional pulses of fluid shear stress, wherein percent viability is determined as the percent viability relative to a control maintained without the application of shear stresses for the duration of the pulse applications.

### Claim 28 (depends on 19)

The method of claim 19, wherein the initial and additional pulses of fluid shear stress apply a fluid shear stress in the range from about 500 dyn/cm2 to about 6500 dyn/cm2.

### Claim 29 (depends on 19)

The method of claim 19, wherein the additional pulses apply a supra-physiologic level of fluid shear stress.

### Claim 30 (depends on 19)

The method of claim 19, wherein the steps of applying the initial pulse of fluid shear stress to the sample and applying one or more additional pulses of fluid shear stress to the sample comprise passing the sample through a conduit at a substantially constant flow rate.

### Claim 31 (depends on 19)

The method of claim 19, wherein the initial pulse and the one or more additional pulses each have a duration of no greater than about 20 msec.

### Claim 32 (depends on 19)

The method of claim 19, wherein at least two additional pulses of fluid shear stress are applied to the sample.

### Claim 33 (depends on 19)

The method of claim 19, further comprising subsequently measuring the concentration of the viable cancerous cells in the sample.

### Claim 34 (depends on 19)

The method of claim 19, further comprising subsequently conducting an assay on the sample, the assay providing a cancer diagnosis or a cancer prognosis for the mammalian subject.