Isolation, cultivation and uses of stem/progenitor cells

• US 9,737,568 B2
• Filed: 05/18/2015
• Issued: 08/22/2017
• 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:
  US14/715,441
• Publication Number:
  US 20160024466A1
• Time in Patent Office: 827 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 of cultivating stem/progenitor cells of the amniotic membrane of umbilical cord that comprises obtaining a tissue explant from the amniotic membrane of umbilical cord and cultivating the tissue explant in suitable cultivation media and cultivation conditions over a suitable period of time. The method may include isolating the stem/progenitor cells from the tissue cultures. The stem cells may be epithelial or mesenchymal stem/progenitor cells. The isolated stem cell cells can have embryonic stem cell-like properties and can be used for various therapeutic purposes.

Claims

1. A method of cultivating a mesenchymal stem/progenitor cell population of the amniotic membrane of umbilical cord, the mesenchymal stem/progenitor cell population having the capacity to differentiate in multiple cell types, the method comprising:

obtaining a tissue explant from the amniotic membrane of umbilical cord;

cultivating the tissue explant under conditions allowing cell proliferation of the mesenchymal stem/progenitor cells without differentiation of the mesenchymal stem/progenitor cells; and

isolating the stem/progenitor cells of the amniotic membrane, 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 secrete Activin A and follistatin.

2. The method of claim 1, further comprising preserving the isolated mesenchymal stem/progenitor cells.

3. The method of claim 2, wherein preserving is carried out by using cryo-preservation.

4. The method of claim 1, wherein the tissue explant is obtained by separating the amniotic membrane from the other components of the umbilical cord in vitro.