Cultures of human CNS neural stem cells

• US 6,103,530 A
• Filed: 10/23/1998
• Issued: 08/15/2000
• Est. Priority Date: 09/05/1997
• Status: Expired due to Term

• Current Assignee: BOCO Silicon Valley, Inc.
• Original Assignee: Cytotherapeutics, Inc.
• Inventors: Carpenter, Melissa
• Primary Examiner(s): Lankford, Jr., Leon B.
• Assistant Examiner(s): TATE, CHRISTOPHER ROBIN
• Application Number:
  US09/178,035
• Time in Patent Office: 662 Days
• Field of Search: 435/325, 435/368, 435/377, 435/384, 435/387, 435/389, 435/404, 435/405, 435/406
• US Class Current: 435/405
• CPC Class Codes: A61K 2035/126 Immunoprotecting barriers, e.g. jackets, diffusion chambers
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61K 35/30 Nerves; Brain; Eyes; Corneal cells; Cerebrospinal fluid; Neuronal stem cells; Neuronal precursor cells; Glial cells; Oligodendrocytes; Schwann cells; Astroglia; Astrocytes; Choroid plexus; Spinal cord tissue
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: C12N 2500/25 Insulin-transferrin; Insulin-transferrin-selenium
• CPC Class Codes: C12N 2500/34 Sugars
• CPC Class Codes: C12N 2500/90 Serum-free medium, which may still contain naturally-sourced components
• CPC Class Codes: C12N 2501/105 Insulin-like growth factors [IGF]
• CPC Class Codes: C12N 2501/11 Epidermal growth factor [EGF]
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2501/235 Leukemia inhibitory factor [LIF]
• CPC Class Codes: C12N 2501/392 Sexual steroids
• CPC Class Codes: C12N 2503/02 Drug screening
• CPC Class Codes: C12N 5/0623 Stem cells
• CPC Class Codes: G01N 33/5005 involving human or animal cells immunoassay G01N33/56966; immunoassays of protozoa G01N33/56905; protozoa in screening assays C12Q1/025

Abstract

Isolation, characterization, proliferation, differentiation and transplantation of mammalian neural stem cells is disclosed.

Claims

1. A culture medium for proliferating mammalian neural stem cells, the media comprising cell viability and cell proliferation effective amounts of the following components:

(a) a standard defined basal medium;

(b) a supplemental carbohydrate, wherein the carbohydrate is present at a concentration between 0.2% and 1.0% weight/volume (w/v);

(c) a supplemental buffer, such that the pH of the culture medium is between pH 6 and pH 8;

(d) supplemental agents, wherein the agents are selected from the group consisting of insulin, transferrin, progesterone, selenium, putrescine, and combinations thereof;

(e) one or more growth factors that stimulate proliferation of neural stem cells, wherein the growth factors are present at a concentration of between 0.2 ng/ml and 200 ng/ml, and;

(f) leukocyte inhibitory factor (LIF), wherein the LIF is present at a concentration of between 0.1 ng/ml and 500 ng/ml.

2. The medium of claim 1 further comprising heparin.