# US 2023/0407263 A1 · Generation of a mesenchymal stromal cell bank from the pooled mononuclear cells of multiple bone marrow donors

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- Number: US 2023/0407263 A1
- Document ID: US20230407263A1
- Status: Published application
- Kind: A1
- Application number: 18/104,924
- Filed: 2023-02-02
- Published: 2023-12-21
- Earliest claimed: 2014-07-16
- Assignees: Johann Wolfgang Goethe-Universität Frankfurt am Main; Drk Blutspendedienst Baden-WüRttemberg-Hessen Ggmbh
- Applicants: Johann Wolfgang Goethe-Universität, Frankfurt Am Main; Drk Blutspendedienst Baden-WüRttemberg-Hessen Ggmbh
- Inventors: Peter Bader; Selim Kuci; Zyrafete Kuci; Halvard Bönig
- Web page: https://patentgrep.com/patent/US20230407263A1

## CPC

- C12N 5/0663: {Bone marrow mesenchymal stem cells (BM-MSC)}
- A61K 35/28: Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
- A61P 1/00: Drugs for disorders of the alimentary tract or the digestive system
- A61P 1/04: for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
- A61P 3/10: for hyperglycaemia, e.g. antidiabetics
- A61P 5/14: of the thyroid hormones, e.g. T3, T4
- A61P 13/12: of the kidneys
- A61P 19/02: for joint disorders, e.g. arthritis, arthrosis
- A61P 25/00: Drugs for disorders of the nervous system
- A61P 27/02: Ophthalmic agents
- A61P 29/00: Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
- A61P 37/02: Immunomodulators
- A61P 37/06: Immunosuppressants, e.g. drugs for graft rejection
- C12Q 1/6881: for tissue or cell typing, e.g. human leukocyte antigen [HLA] probes
- A61K 2035/124: {the cells being hematopoietic, bone marrow derived or blood cells}
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES

## Abstract

The present invention pertains to an improved mesenchymal stromal cell (MSC) preparation and a method for producing the same. The invention provides a new strategy to isolate MSC from bone marrow mononuclear cells (BM-MNCs) by pooling BM-MNCs of multiple unrelated (third-party) bone marrow donors. The MSC preparation manufactured in accordance with the methodology of the invention is characterized by a stable proliferative capability and an increased immunosuppressive potential when compared to individual donor MSC preparations or a pool of individual MSCs generated from multiple donors. The MSCs prepared according to the invention are particularly useful for medical applications such as the treatment of graft-versus-host disease (GvHD) in recipients with hematopoietic stem cell transplants, patients with autoimmune disorders and as a cell-based therapy in regenerative medicine.

## Claims

### Claim at position 1

-15. (canceled)

### Claim 16

A mesenchymal stromal cells (MSC) preparation obtainable by a method comprising the steps of:
a) pooling individual bone marrow samples obtained from at least two genetically distinct donors to obtain a sample cell-pool; and
b) isolating mesenchymal stromal cells from said sample cell pool.

### Claim 17 (depends on 16)

The MSC preparation of to claim 16, wherein the method further incudes the step of culturing said sample cell-pool prior to isolating mesenchymal stromal cells from said sample cell-pool.

### Claim 18 (depends on 16)

The MSC preparation of claim 16, wherein said bone marrow sample is a mammalian bone marrow sample and said mesenchymal stromal cell is a mammalian mesenchymal stromal cell.

### Claim 19 (depends on 18)

The MSC preparation of claim 18, wherein the mammalian bone marrow sample is a human bone marrow sample and the mammalian mesenchymal stromal cell is a human mesenchymal stromal cell.

### Claim 20 (depends on 16)

The MSC preparation of to claim 16, wherein said bone marrow samples are bone marrow mononuclear cell samples.

### Claim 21 (depends on 16)

The MSC preparation of to claim 16, wherein said bone marrow samples are obtained from at least three, at least four, at least five, at least six, at least seven, or at least eight genetically distinct donors.

### Claim 22 (depends on 16)

The MSC preparation of to claim 16, wherein the method further includes the step of storing said isolated mesenchymal stromal cells.

### Claim 23 (depends on 16)

The MSC preparation of claim 16, further including the steps of:
a) determining the hTERT negative and polygenic status of the isolated mesenchymal stromal cells; and
b) enriching the isolated mesenchymal stromal cells to obtain hTERT negative and polygenic isolated mesenchymal stromal cells.

### Claim 24 (depends on 23)

The MSC preparation of claim 23, wherein the method further includes the step of enriching the hTERT and polygenic isolated, enriched mesenchymal stromal cells to:
(a) At least 95% CD73+ cells, and/or
(b) At least 95% CD90+ cells, and/or
(c) At least 95% CD105+ cells, and/or
(d) At least 95% HLA-class I+ cells, and/or
(e) Less than 1% CD45+ cells, and/or
(f) Less than 1% CD14+ cells, and/or
(g) Less than 1% CD34+ cells, and/or
(h) Less than 5% HLA-DR+ cells.

### Claim 25 (depends on 24)

The MSC preparation of claim 24, wherein the hTERT and polygenic isolated mesenchymal stromal cells are (a), (b) and (c), and/or (e), (f) and (g).

### Claim 26 (depends on 16)

The MSC preparation of claim 16, wherein the method further includes the step of culturing the isolated mesenchymal stromal cells.

### Claim 27 (depends on 16)

The MSC preparation of claim 16, wherein the method furthermore includes expanding the isolated mesenchymal stromal cells until receiving an amount of mesenchymal stromal cells applicable in the clinic.

### Claim 28 (depends on 27)

The MSC preparation of claim 27, wherein the isolated mesenchymal stromal cells are expanded immediately after isolating the mesenchymal stromal cells, or wherein the isolated mesenchymal stromal cells are expanded from a cryopreserved aliquot of the isolated mesenchymal stromal cells.

### Claim 29

An isolated cellular composition comprising bone marrow samples from at least two genetically distinct bone marrow donors, wherein each of said bone marrow samples comprises at least one non-adherent cell fraction and at least one adherent cell fraction.

### Claim 30 (depends on 30)

The isolated cellular composition of to claim 30, wherein each of said bone marrow samples comprises a bone marrow mononuclear cell fraction.

### Claim 31

A method for the treatment of a disease in a subject in need of the treatment, wherein the treatment comprises administering a therapeutically effective amount of an MSC-preparation obtainable by a method comprising the steps of:
a) pooling individual bone marrow samples obtained from at least two genetically distinct donors to obtain a sample cell-pool; and
b) isolating mesenchymal stromal cells from said sample cell pool.

### Claim 32 (depends on 31)

The method of claim 31, wherein the disease is an autoimmune disease selected from the group consisting of multiple sclerosis, Type 1 diabetes, rheumatoid arthritis, uveitis, autoimmune thyroid disease, inflammatory bowel disease (IBD), scleroderma, Graves' disease, lupus, Crohn's disease, autoimmune lymphoproliferative disease (ALPS), demyelinating disease, autoimmune encephalomyelitis, autoimmune gastritis (AIG), autoimmune glomerular diseases, and graft-versus-host disease (GvHD).