Isolation, cultivation and uses of stem/progenitor cells

• US 10,363,275 B2
• Filed: 08/15/2005
• Issued: 07/30/2019
• Est. Priority Date: 08/16/2004
• Status: Active Grant

• Current Assignee: CellResearch Corporation Pte Ltd (CellResearch Group of Cos.)
• Original Assignee: CellResearch Corporation Pte Ltd (CellResearch Group of Cos.)
• Inventors: Phan, Toan-Thang, Lim, Ivor Jiun
• Primary Examiner(s): Kim, Taeyoon
• Application Number:
  US11/205,248
• Publication Number:
  US 20060078993A1
• Time in Patent Office: 5,097 Days
• CPC Class Codes: A61K 35/28 Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
• CPC Class Codes: A61K 35/36 Skin; Hair; Nails; Sebaceous glands; Cerumen; Epidermis; Epithelial cells; Keratinocytes; Langerhans cells; Ectodermal cells islets of Langerhans A61K35/39
• CPC Class Codes: A61K 35/51 Umbilical cord; Umbilical cord blood; Umbilical stem cells
• CPC Class Codes: A61K 9/0014 Skin, i.e. galenical aspects of topical compositions non-active ingredients are additionally classified in A61K47/00; A61K9/0009, A61K9/0021, A61K9/7015, A61K9/7023 take precedence; cosmetic preparations A61K8/00, A61Q; preparations for wound dressings or bandages A61L26/00
• CPC Class Codes: A61K 9/06 Ointments; Bases therefor; Other semi-solid forms, e.g. creams, sticks, gels composition of ointments, creams or gels A61K47/00
• CPC Class Codes: A61P 1/18 for pancreatic disorders, e.g. pancreatic enzymes
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 17/16 Emollients or protectives, e.g. against radiation
• CPC Class Codes: A61P 17/18 Antioxidants, e.g. antiradicals preparations for protection against sunlight A61Q17/00
• CPC Class Codes: A61P 25/14 for treating abnormal movements, e.g. chorea, dyskinesia
• CPC Class Codes: A61P 25/16 Anti-Parkinson drugs
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 5/14 of the thyroid hormones, e.g. T3, T4
• CPC Class Codes: A61P 5/24 of the sex hormones
• CPC Class Codes: A61P 5/38 of the suprarenal hormones
• CPC Class Codes: A61P 5/48 of the pancreatic hormones
• CPC Class Codes: A61P 7/06 Antianaemics
• CPC Class Codes: C12N 2506/025 : from extra-embryonic cells, e.g. trophoblast, placenta
• CPC Class Codes: C12N 2506/1392 : from mesenchymal stem cells from other natural sources
• CPC Class Codes: C12N 5/0603 : Embryonic cells production of embryos, nuclear transfer A01K67/027; Embryoid bodies
• CPC Class Codes: C12N 5/0605 : Cells from extra-embryonic tissues, e.g. placenta, amnion, yolk sac, Wharton's jelly
• CPC Class Codes: C12N 5/0606 : Pluripotent embryonic cells, e.g. embryonic stem cells [ES] embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696
• CPC Class Codes: C12N 5/0629 : Keratinocytes; Whole skin
• CPC Class Codes: C12N 5/063 : Kereatinocyte stem cells; Keratinocyte progenitors
• CPC Class Codes: C12N 5/0668 : Mesenchymal stem cells from other natural sources

Abstract

The present invention relates to a method for isolating stem/progenitor cells from the amniotic membrane of umbilical cord, wherein the method comprises separating the amniotic membrane from the other components of the umbilical cord in vitro, culturing the amniotic membrane tissue under conditions allowing cell proliferation, and isolating the stem/progenitor cells from the tissue cultures. The isolated stem cell cells can have embryonic stem cell-like properties and can be used for various therapeutic purposes. In one embodiment, the invention relates to the isolation and cultivation of stem cells such as epithelial and/or mesenchymal stem/progenitor cells under conditions allowing the cells to undergo mitotic expansion. Furthermore, the invention is directed to a method for the differentiation of the isolated stem/progenitor cells into epithelial and/or mesenchymal cells.

Claims

1. A mesenchymal stem/progenitor cell population isolated from the amniotic membrane of umbilical cord, the mesenchymal stem/progenitor cell population having the capacity to differentiate into multiple cell types and being isolated by a method comprising:

(a) separating the amniotic membrane from the other components of the umbilical cord in vitro;

(b) culturing the amniotic membrane tissue obtained in (a) as a tissue explant, or culturing amniotic membrane cells separated from the amniotic membrane tissue obtained in (a), under conditions allowing cell proliferation of mesenchymal stem/progenitor cells without differentiation of the mesenchymal stem/progenitor cells; and

(c) harvesting the mesenchymal stem/progenitor cells, wherein the stem/progenitor cells have a spindle shape, express the following genes;

POU5f1, Bmi-1, leukemia inhibitory factor (LIF), and wherein the stem/progenitor cells secrets Activin A and Follistatin.

2. The stem/progenitor cell of claim 1 isolated from the amniotic membrane of umbilical cord, wherein the amniotic membrane cells are separated from the amniotic membrane tissue obtained in (a) by enzymatic separation.

3. The stem/progenitor cell of claim 1, wherein the isolation further comprises

(d) culturing the stem/progenitor cells under conditions allowing the cells to undergo clonal expansion.

4. The stem/progenitor cell of claim 1, wherein the isolation further comprises preserving the isolated stem/progenitor cells for further use.

5. The stem/progenitor cell of claim 4, wherein preserving is carried out by using cryo-preservation.

6. A stem/progenitor cell of claim 1, wherein the stem/progenitor cell express the following growth factors:

connective tissue growth factor (CTGF), vascular endothelial growth factor (VEGF), placenta-like growth factor PLGF, STAT3, stem cell factor (SCF), Hepatoma-derived Growth Factor (HDGF), Fibroblast Growth Factor-2 (FGF-2), Platelet-derived Growth Factor (PDGF), alpha-Smooth Muscle Actin (α

-SMA), Fibronectin, Decorin, and Syndecan-1,2,3,4.

7. A pharmaceutical composition comprising a member selected from the group consisting of a stem/progenitor cell as defined in claim 1 isolated from the amniotic membrane of umbilical cord and a cellular extract thereof.

8. The pharmaceutical composition of claim 7, wherein the pharmaceutical composition is adapted for systemic or topical application.

9. The pharmaceutical composition of claim 8, wherein the pharmaceutical composition adapted for topical application is selected from the group consisting of an ointment, a cream, and a lotion.