# US 11,028,362 B2 · Decellularized human amniotic membrane for cell delivery, cell culture and inflammation prevention

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- Number: US 11,028,362 B2
- Document ID: US11028362B2
- Status: Granted
- Kind: B2
- Application number: 15/567,058
- Filed: 2016-04-15
- Published: 2018-04-12
- Granted: 2021-06-08
- Earliest claimed: 2015-04-17
- Assignees: The Regents of the University of California
- Applicants: The Regents of the University of California
- Inventors: Volodymyr Ryzhuk; Aijun Wang; Diana Farmer; Benjamin Keller
- Web page: https://patentgrep.com/patent/US11028362B2

## CPC

- C12N 5/0605: {Cells from extra-embryonic tissues, e.g. placenta, amnion, yolk sac, Wharton's jelly}
- A61K 9/0019: {Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner (non-active ingredients are additionally classified in A61K47/00)}
- A61K 35/28: Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
- A61K 35/39: Pancreas; Islets of Langerhans (Langerhans cells of epidermis A61K35/36)
- A61K 35/50: Placenta; Placental stem cells; Amniotic fluid; Amnion; Amniotic stem cells
- A61K 38/488: {Aspartic endopeptidases (3.4.23), e.g. pepsin, chymosin, renin, cathepsin E}
- A61L 26/0057: {Ingredients of undetermined constitution or reaction products thereof}
- A61L 26/008: {Hydrogels or hydrocolloids}
- A61L 27/3604: {characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel}
- A61L 27/3683: {subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment}
- A61L 27/3834: {Cells able to produce different cell types, e.g. hematopoietic stem cells, mesenchymal stem cells, marrow stromal cells, embryonic stem cells}
- A61L 27/52: Hydrogels or hydrocolloids
- A61P 17/02: for treating wounds, ulcers, burns, scars, keloids, or the like
- A61P 19/08: for bone diseases, e.g. rachitism, Paget's disease
- A61P 29/00: Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
- C12N 5/063: {Kereatinocyte stem cells; Keratinocyte progenitors}
- C12N 5/0662: {Stem cells}
- C12N 5/0668: {Mesenchymal stem cells from other natural sources}
- A61L 2430/38: for reconstruction of the spine, vertebrae or intervertebral discs
- C12N 2533/54: Collagen; Gelatin
- C12N 2533/92: Amnion; Decellularised dermis or mucosa
- 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)
- A61L: METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES (preservation of bodies or disinfecting characterised by the agents employed A01N; preserving, e.g. sterilising, food or foodstuffs A23; preparations for medical, dental or toiletry purposes A61K)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS

## Abstract

Provided herein is an isolated decelluarized amniotic membrane (DCM) and methods for using same therapeutically in vivo and in vitro. In one aspect, the isolated DCM is further processed, freezing, freeze drying, lyophilization micronized into powder or treatment with pepsin to create a hydrogel.

## Claims

### Claim 1

A hydrogel composition comprising:
a powdered and freeze dried decellularized amniotic membrane (DCM), the amniotic membrane being decellularized using a method comprising:
adding to the amniotic membrane about a 3% detergent solution comprising t-octylphenoxypolyethoxyethanol solution to lyse the cells,
rinsing with de-ionized water to remove the detergent solution,
adding about a 4% sodium deoxycholic acid solution to further decellularize the membrane,
rinsing with de-ionized water to remove the acid solution,
adding a solution of about 0.1% peracetic acid/4% ethanol solution to remove residual nucleic acids,
rinsing with a composition comprising phosphate buffered saline and subsequently de-ionized water, and
removing excess water;
stem cells;
a non-cytotoxic cross-linking agent; and
a carrier comprising buffered saline.

### Claim 2 (depends on 1)

The hydrogel composition of claim 1, wherein the DCM is processed by lyophilization and micronized into powder.

### Claim 3 (depends on 1)

The hydrogel composition of claim 1, wherein the DCM is derived from placenta isolated from a human, an ovine, a porcine, a bovine, an equine, a feline, a canine or a murine.

### Claim 4 (depends on 1)

The hydrogel composition of claim 1, wherein the carrier is a pharmaceutically acceptable carrier comprising buffered saline.

### Claim 5 (depends on 4)

The hydrogel composition of claim 4, wherein the pharmaceutically acceptable carrier further comprises deionized water or a collagen-based product.

### Claim 6 (depends on 1)

The hydrogel composition of claim 1, wherein the stem cells comprise mesenchymal stem cells, bone marrow stem cells, or placental derived mesenchymal stem cells (PMSCs).

### Claim 7 (depends on 1)

The hydrogel composition of claim 1, further comprising pepsin.

### Claim 8 (depends on 1)

The hydrogel composition of claim 1, wherein the hydrogel composition is a topical gel.

### Claim 9 (depends on 1)

The hydrogel composition of claim 1, further comprising one or more antibiotics, anti-inflammatory agents or growth factors.

### Claim 10 (depends on 1)

A kit comprising the hydrogel composition of claim 1.

### Claim 11 (depends on 10)

The kit of claim 10, further comprising instructions for use.

### Claim 12 (depends on 10)

The kit of claim 10, further comprising instructions for administration by one or more of topical, intraperitoneal, implantation, local, or systemic administration route.

### Claim 13 (depends on 1)

A method to culture a cell, comprising mixing an exogenous cell with the hydrogel composition of claim 1, and culturing the exogenous cell under conditions that favor growth and expansion of the exogenous cell.

### Claim 14 (depends on 13)

The method of claim 13, wherein the mixing is in vitro or in vivo.

### Claim 15 (depends on 13)

The method of claim 13, wherein the exogenous cell is selected from the group consisting of a stem cell and a mesenchymal stem cell.

### Claim 16 (depends on 1)

A method to treat a wound, comprising administering to a subject in need thereof, an effective amount of the hydrogel composition of claim 1, thereby treating the wound.

### Claim 17 (depends on 1)

A method for regulating, treating or preventing inflammation in a subject in need thereof, comprising administering to the subject in need thereof, an effective amount of the hydrogel composition of claim 1, thereby regulating, treating or preventing inflammation.

### Claim 18 (depends on 1)

A method for treating or preventing adhesion or scar formation in a subject in need thereof, comprising administering to the subject in need thereof, an effective amount of the hydrogel composition of claim 1, thereby treating or preventing adhesion or scar formation.

### Claim 19 (depends on 1)

A method to treat spinal cord injury or Spina bifida, comprising administering to a subject in need thereof an effective amount of the hydrogel composition of claim 1, thereby treating spinal cord injury or Spina bifida.

### Claim 20 (depends on 1)

A method to promote vascularization in a subject in need thereof, comprising administering to the subject in need thereof, an effective amount of the hydrogel composition of claim 1, thereby promoting vascularization in the subject.

### Claim 21 (depends on 16, 17, 18)

The method of any one of claims 16 to 18, wherein administration comprises one or more of topical, intraperitoneal, implantation, local, or systemic.

### Claim 22 (depends on 21)

The method of claim 21, wherein the stem cells are autologous or allogeneic to the subject being treated.

### Claim 23 (depends on 16, 17, 18, 19, 20)

The method of any one of claims 16 to 20, wherein the subject is a human, a bovine, an equine, a feline, a canine or a murine.

### Claim 24 (depends on 16, 17, 18, 19, 20)

The method of any one of claims 16 to 20, wherein the composition further comprises one or more antibiotics, anti-inflammatory agents or growth factors.

### Claim 25 (depends on 1)

A method to promote neural tissue repair or regeneration, comprising administering to a subject in need thereof an effective amount of the hydrogel composition of claim 1, thereby promoting neural tissue repair or regeneration.