# US 12,180,447 B2 · Bone marrow microfluidic devices and methods for preparing and using the same

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- Number: US 12,180,447 B2
- Document ID: US12180447B2
- Status: Granted
- Kind: B2
- Application number: 18/343,514
- Filed: 2023-06-28
- Published: 2023-11-09
- Granted: 2024-12-31
- Earliest claimed: 2017-03-24
- Assignees: President and Fellows of Harvard College; The General Hospital Corporation
- Applicants: President and Fellows of Harvard College; The General Hospital Corporation
- Inventors: David Benson Chou; Liliana S. Teixeira Moreira Leijten; Arianna Rech; Richard Novak; Donald E. Ingber; Yuka Milton; Viktoras Frismantas; Oren Levy
- Web page: https://patentgrep.com/patent/US12180447B2

## CPC

- C12M 23/16: {Microfluidic devices; Capillary tubes (integrated microfluidic structures B01L3/5027; microreactors B01J19/0093)}
- B01L 3/502715: {characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces}
- C12M 23/34: {Internal compartments or partitions}
- C12M 25/02: {Membranes; Filters (filters or filtration in general B01D24/00-B01D41/00)}
- C12M 25/14: {Scaffolds; Matrices (in general C12N5/0068)}
- C12M 29/04: {Filters; Permeable or porous membranes or plates, e.g. dialysis}
- C12M 29/10: {Perfusion}
- C12M 35/08: {Chemical, biochemical or biological means, e.g. plasma jet, co-culture}
- C12N 5/0068: {General culture methods using substrates (for specific animal cell type C12N5/06)}
- C12N 5/0669: {Bone marrow stromal cells; Whole bone marrow (isolated stem cells from bone marrow C12N5/0647, C12N5/0663)}
- G01N 33/5008: {for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics}
- B01L 2300/12: Specific details about materials
- C12N 2501/40: Regulators of development
- C12N 2501/999: Small molecules not provided for elsewhere
- C12N 2502/28: Vascular endothelial cells
- C12N 2521/00: Culture process characterised by the use of hydrostatic pressure, flow or shear forces
- C12N 2533/50: Proteins
- C12N 2533/54: Collagen; Gelatin
- C12N 2533/56: Fibrin; Thrombin
- C12N 2533/70: Polysaccharides
- C12N 2533/90: Substrates of biological origin, e.g. extracellular matrix, decellularised tissue
- C12M: APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; {APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS}
- 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)
- 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

The present disclosure relates to a microfluidic devices and methods for culturing bone marrow cells. Aspects include methods of preparing microfluidic devices and culturing bone marrow cells with the microfluidic devices. In some aspects, a method includes providing a microfluidic device having an upper chamber, a lower chamber, and a porous membrane separating the upper chamber from the lower chamber. The method further includes seeding walls of the lower chamber and a bottom surface of the membrane with endothelial cells. The method further includes providing a matrix within the upper chamber. The matrix includes fibrin gel and bone marrow cells. The method further includes filling or perfusing the upper chamber with a media.

## Claims

### Claim 1

A method of preparing a microfluidic device, the method comprising:
providing a microfluidic device having a first microchannel, a second microchannel, and a porous membrane separating the first microchannel from the second microchannel;
providing a matrix within the first microchannel, the matrix comprising CD34+ bone marrow progenitor cells;
perfusing media through the second microchannel;
producing cells differentiated from said CD34+ bone marrow progenitor cells, said differentiated cells comprising erythroid lineage cells.

### Claim 2 (depends on 1)

The method of claim 1, wherein the matrix is a fibrin gel.

### Claim 3 (depends on 2)

The method of claim 2, wherein said fibrin gel comprises aprotinin.

### Claim 4 (depends on 1)

The method of claim 1, further comprising seeding the second microchannel with endothelial cells.

### Claim 5 (depends on 1)

The method of claim 1, wherein the first microchannel forms a closed channel.

### Claim 6 (depends on 1)

The method of claim 1, wherein said matrix further comprises stromal cells.

### Claim 7 (depends on 1)

The method of claim 1, wherein there is no flow of media through said first microchannel.

### Claim 8 (depends on 7)

The method of claim 7, wherein media in said second microchannel can migrate through said membrane into said first microchannel.