# US 2020/0399610 A1 · Materials and methods for generating therapeutic mesenchymal stem cells

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- Number: US 2020/0399610 A1
- Document ID: US20200399610A1
- Status: Published application
- Kind: A1
- Application number: 16/647,345
- Filed: 2018-09-14
- Published: 2020-12-24
- Earliest claimed: 2017-09-14
- Assignees: University of Kansas
- Applicants: University of Kansas
- Inventors: Hiroshi Nishimune; Richard Barohn; Buddhadeb Dawn; Yomna Badawi; James W Mitchell; Rupal Soder
- Web page: https://patentgrep.com/patent/US20200399610A1

## CPC

- A61K 35/28: Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
- C12N 5/0668: {Mesenchymal stem cells from other natural sources}
- A61K 35/51: Umbilical cord; Umbilical cord blood; Umbilical stem cells
- C12N 5/0619: {Neurons}
- A61K 38/00: Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00)
- C12N 2500/32: Amino acids
- C12N 2500/99: Serum-free medium
- C12N 2501/10: Growth factors
- C12N 2501/105: Insulin-like growth factors [IGF]
- C12N 2501/11: Epidermal growth factor [EGF]
- C12N 2501/115: Basic fibroblast growth factor (bFGF, FGF-2)
- C12N 2501/13: Nerve growth factor [NGF]; Brain-derived neurotrophic factor [BDNF]; Cilliary neurotrophic factor [CNTF]; Glial-derived neurotrophic factor [GDNF]; Neurotrophins [NT]; Neuregulins
- C12N 2501/135: Platelet-derived growth factor [PDGF]
- C12N 2501/165: Vascular endothelial growth factor [VEGF]
- C12N 2501/195: Heregulin, neu differentiation factor
- C12N 2502/1388: Mesenchymal stem cells from other natural sources
- C12N 2533/52: Fibronectin; Laminin
- 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)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

Embodiments of the present disclosure relate generally to the production of therapeutic mesenchymal stem cells (MSCs). More particularly, the present disclosure relates to the use of cell culture compositions and methods for generating MSCs that secrete neurotrophic factors and synaptic organizing agents for the treatment of neurodegenerative diseases such as Amyotrophic Lateral Sclerosis (ALS). As such, the present disclosure addresses the need for establishing a reliable source of therapeutic stem cells useful for the treatment of neurodegenerative diseases.

## Claims

### Claim 1

An isolated non-genetically modified human cell activated ex vivo from a mesenchymal stem cell (MSC) under conditions such that the isolated non-genetically modified human cell secretes laminin β2 at a level that is greater than the basal secretion level of laminin β2 by the MSC.

### Claim 2 (depends on 1)

The isolated human cell of claim 1, wherein the MSC is obtained from Wharton's jelly from an umbilical cord.

### Claim 3 (depends on 1)

The isolated human cell of claim 1, wherein the MSC is CD140b positive.

### Claim 4 (depends on 1)

The isolated human cell of claim 1, wherein the cell is activated in a cell culture composition comprising cell culture media and Insulin-like growth factor 1 (IGF-1)

### Claim 5 (depends on 4)

The isolated human cell of claim 4, wherein the cell culture composition further comprises one or more additional growth factors selected from Fibroblast Growth Factor (FGF) and Platelet-derived Growth Factor (PDGF).

### Claim 6 (depends on 4)

The isolated human cell of claim 4, wherein the cell culture composition further comprises Heregulin β1.

### Claim 7 (depends on 4)

The isolated human cell of claim 4, wherein the cell culture media is DMEM-F12 containing L-glutamine.

### Claim 8 (depends on 4)

The isolated human cell of claim 4, wherein the cell culture composition further comprises dibutryl cAMP.

### Claim 9 (depends on 4)

The isolated human cell of claim 4, wherein the cell culture composition further comprises 3-isobutyl-1-methylxanthine (IBMX).

### Claim 10 (depends on 1)

The isolated human cell of claim 1, wherein the isolated non-genetically modified human cell further secretes one or more of glial cell-derived neurotrophic factor (GDNF), brain-derived neurotrophic factor (BDNF), and vascular endothelial growth factor (VEGF) at a level that is greater than the basal secretion level of GDNF, BDNF, or VEGF by the MSC.

### Claim 11 (depends on 1)

The isolated human cell of claim 1, wherein the isolated human cell enhances growth and/or survival of one or more motor nerve terminals at a neuromuscular junction upon exposure to the activated MSCs.

### Claim 12 (depends on 1)

The isolated human cell of claim 1, wherein the isolated human cell ameliorates denervation at a neuromuscular junction caused by Amyotrophic Lateral Sclerosis (ALS).

### Claim 13 (depends on 1)

A method of treating a disease for which administration of neurotrophic factors is beneficial in a subject in need thereof, comprising administering to the subject the isolated human cell of claim 1.

### Claim 14 (depends on 13)

The method of claim 13, wherein the diseases is ALS.

### Claim 15

A method for producing a cell from a mesenchymal stem cell (MSC), such that the cell secretes laminin β2 at a level that is greater than the basal secretion level of laminin β2 by the MSC, the method comprising:
exposing the MSC to a cell culture composition comprising cell culture media and Insulin-like growth factor 1 (IGF-1).

### Claim 16 (depends on 15)

The method of claim 15, wherein the MSC is obtained from Wharton's jelly from an umbilical cord.

### Claim 17 (depends on 15)

The method of claim 15, wherein the MSC is CD140b positive.

### Claim 18 (depends on 15)

The method of claim 15, wherein the cell culture composition further comprises one or more additional growth factors selected from Fibroblast Growth Factor (FGF) and Platelet-derived Growth Factor (PDGF).

### Claim 19 (depends on 15)

The method of claim 15, wherein the cell culture composition further comprises Heregulin β1.

### Claim 20 (depends on 15)

The method of claim 15, wherein the cell culture media is DMEM-F12 containing L-glutamine.

### Claim 21 (depends on 15)

The method of claim 15, wherein the cell culture composition further comprises dibutryl cAMP.

### Claim 22 (depends on 15)

The method of claim 15, wherein the cell culture composition further comprises 3-isobutyl-1-methylxanthine (IBMX).

### Claim 23 (depends on 15)

The method of claim 15, wherein the cell further secretes one or more of glial cell-derived neurotrophic factor (GDNF), brain-derived neurotrophic factor (BDNF), and vascular endothelial growth factor (VEGF) at a level that is greater than the basal secretion level of GDNF, BDNF, or VEGF by the MSC.

### Claim 24 (depends on 15)

The method of claim 15, wherein the cell enhances growth and/or survival of one or more motor nerve terminals at a neuromuscular junction upon exposure to the cell.

### Claim 25 (depends on 15)

The method of claim 15, wherein the cell ameliorates denervation at a neuromuscular junction caused by Amyotrophic Lateral Sclerosis (ALS).