Dopaminergic neurons derived from corneal limbus, methods of isolation and uses thereof

• US 8,187,875 B2
• Filed: 08/10/2006
• Issued: 05/29/2012
• Est. Priority Date: 02/26/2004
• Status: Expired due to Fees

• Current Assignee: Reliance Life Sciences Pvt. Ltd.
• Original Assignee: Reliance Life Sciences Pvt. Ltd.
• Inventors: Ravindran, Geeta, Rao, Harinarayana
• Primary Examiner(s): LI, QIAN JANICE
• Application Number:
  US11/502,338
• Publication Number:
  US 20070020247A1
• Time in Patent Office: 2,119 Days
• Field of Search: None
• US Class Current: 435/371
• 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: C12N 2500/25 Insulin-transferrin; Insulin-transferrin-selenium
• CPC Class Codes: C12N 2500/30 Organic components
• CPC Class Codes: C12N 2500/62 DMSO
• CPC Class Codes: C12N 2501/01 Modulators of cAMP or cGMP, e.g. non-hydrolysable analogs, phosphodiesterase inhibitors, cholera toxin
• 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 2501/119 Other fibroblast growth factors, e.g. FGF-4, FGF-8, FGF-10
• CPC Class Codes: C12N 2501/235 Leukemia inhibitory factor [LIF]
• CPC Class Codes: C12N 2506/08 from cells of the nervous system
• CPC Class Codes: C12N 2533/32 Polylysine, polyornithine
• CPC Class Codes: C12N 2533/52 Fibronectin; Laminin
• CPC Class Codes: C12N 2533/54 Collagen; Gelatin
• CPC Class Codes: C12N 2533/90 Substrates of biological origin, e.g. extracellular matrix, decellularised tissue
• CPC Class Codes: C12N 5/0607 Non-embryonic pluripotent stem cells, e.g. MASC induced pluripotent stem cells C12N5/0696
• CPC Class Codes: C12N 5/0621 Eye cells, e.g. cornea, iris pigmented cells photoreceptors C12N5/062

Abstract

The present disclosure describes the generation of neural cells and neurons from mammalian pluripotent embryonic-like stem cells (ELSCs) isolated from corneal limbal tissue, a non-embryonic tissue. Specifically, the present disclosure describes the generation of neuroprogenitor cells and differentiated dopaminergic neurons from ELSCs. The disclosed methods demonstrate the potential of ELSCs as a therapeutic tool, and may provide new therapeutic alternatives for various diseases, conditions, and injuries. Neuroprogenitor cells generated from ELSCs isolated from corneo-limbal tissue were transplanted into a rat model of Parkinson'"'"'s disease, and were able to alleviate motor abnormalities in the rats for at least six months.

Claims

1. A population of neural precursor cells in an in vitro culture generated by differentiating human pluripotent embryonic-like stem cells (ELSCs) derived from human corneoscleral limbus tissue, and isolated based on the expression of SSEA-4, keratin growth factor (“

KGF”

), collagen-I and p63, into neural precursor cells.

2. The cell population of claim 1, wherein the ELSCs:

i) are capable of proliferating in an in vitro culture;

ii) maintain the potential to differentiate into cells of endoderm, mesoderm or ectoderm lineage in culture; and

iii) are capable of forming embryoid-like bodies when placed in suspension culture.

3. The cells of claim 1, wherein said population of neural precursor cells also expresses keratin.