Production from blood of cells of neural lineage

• US 20080318314A1
• Filed: 06/20/2007
• Published: 12/25/2008
• Est. Priority Date: 06/20/2007
• Status: Abandoned Application

• Current Assignee: Kwalata Trading Ltd.
• Original Assignee: Kwalata Trading Ltd.
• Inventors: Porat, Yael, Fulga, Valentin, Porozov, Svetlana, Fisher, Yehudit
• Application Number:
  US11/820,991
• Publication Number:
  US 20080318314A1
• Time in Patent Office: Days
• US Class Current: 435/373
• 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 2501/13 Nerve growth factor [NGF]; Brain-derived neurotrophic factor [BDNF]; Cilliary neurotrophic factor [CNTF]; Glial-derived neurotrophic factor [GDNF]; Neurotrophins [NT]; Neuregulins
• CPC Class Codes: C12N 2501/91 Heparin
• CPC Class Codes: C12N 2506/11 from blood or immune system cells
• CPC Class Codes: C12N 5/0618 Cells of the nervous system

Abstract

A method including in vitro stimulating a core cell population (CCP) of at least 5 million cells that have a density of less than 1.072 g/ml, and at least 1.5% of which are CD34+CD45−/dim, to differentiate into a neural progenitor/precursor cell population (NPCP). Other embodiments are also described.

Claims

1. A method comprising in vitro stimulating a core cell population (CCP) of at least 5 million cells that have a density of less than 1.072 g/ml, and at least 1.5% of which are CD34+CD45−

/dim, to differentiate into a neural progenitor/precursor cell population (NPCP).

3. The method according to claim 1, wherein at least 1.5 million of the cells in the NPCP and at least 1.5% of the cells in the NPCP comprise at least one molecule or molecular structure selected from the list consisting of:

CD34, CD117, CD44, Neu-N, nestin, microtubule associated protein-1 (MAP-1), MAP Tau, microtubule associated protein-2 (MAP-2), neurofilament NF200, neuron-specific enolase (NSE), choline acetyltransferase (ChAT) neuronal class III β

-Tubulin, glutamic acid decarboxylase (GAD), S-100, GDNF, CNTF, BDNF, NGF, NT3, NT4/5, glutamic acid, gamma-aminobutyric acid (GABA), oligodendrocyte marker (O4), myelin basic protein (MBP), galactocerebroside (GalC), glial fibrillary acidic protein (GFAP), CD211b, dopamine, norepinephrine, epinephrine, glutamate, glycine, acetylcholine, serotonin, and endorphin.

4. The method according to claim 1, comprising preparing the NPCP as a research tool.

6. The method according to claim 1, comprising applying cells extracted from a mammalian donor to one or more gradients suitable for selecting cells having a density less than 1.072 g/ml, and deriving the CCP responsive to applying the cells to the gradient.

10. The method according to claim 1, wherein the CCP is characterized by at least 30% of the CCP being CD14+, and wherein stimulating the CCP comprises stimulating the CCP having the at least 30% of the CCP that are CD14+.

12. The method according to claim 1, wherein the CCP is characterized by at least 40% of the CCP being CD31+, and wherein stimulating the CCP comprises stimulating the CCP having the at least 40% of the CCP that are CD31+.

13. The method according to claim 1, wherein stimulating the CCP comprises stimulating the CCP to differentiate into a pre-designated, desired class of neural progenitor cells.

14. The method according to claim 1, wherein stimulating the CCP comprises culturing the CCP during a period of between 3 and 60 days in vitro.

15. The method according to claim 1, comprising deriving the CCP from at least one source selected from the list consisting of:

umbilical cord blood, umbilical cord tissue, neonatal tissue, adult tissue, fat tissue, nervous tissue, bone marrow, mobilized blood, peripheral blood, peripheral blood mononuclear cells, and skin tissue.

16. The method according to claim 1, comprising deriving the CCP from frozen tissue.

18. The method according to claim 1, wherein stimulating the CCP comprises incubating the CCP in a container having a surface comprising at least one of:

an antibody and autologous plasma.

22. The method according to claim 1, wherein stimulating the CCP comprises culturing the CCP in the presence of at least one of the following:

B27, F12, a proliferation-differentiation-enhancing agent, anti-CD34, anti-CD117, LIF, IGF, b-FGF, M-CSF, GM-CSF, TGF alpha, TGF beta, BHA, BDNF, NGF, NT3, NT4/5, S-100, GDNF, CNTF, EGF, NGF3, CFN, ADMIF, estrogen, prolactin, an adrenocorticoid, glutamate, serotonin, acetylcholine, retinoic acid (RA), heparin, insulin, forskolin, cortisone, and a derivative of any of these.

24. The method according to claim 1, comprising freezing the CCP prior to stimulating the CCP.

25. The method according to claim 1, comprising freezing the NPCP.

26. The method according to claim 1, comprising transporting the CCP to a site at least 10 km from a site where the CCP is first created, and stimulating the CCP at the remote site.

27. The method according to claim 1, comprising transporting the NPCP to a site at least 10 km from a site where the NPCP is first created.

34. The method according to claim 1, comprising preparing the NPCP as a product for administration to a patient.

36. The method according to claim 1, comprising facilitating a diagnosis responsive to stimulating the CCP to differentiate into the NPCP.

38. The method according to claim 1, wherein stimulating the CCP comprises incubating the CCP in a container with a surface comprising a growth-enhancing molecule other than collagen or fibronectin.

39. The method according to claim 38, wherein incubating the CCP comprises incubating the CCP in a container having a surface that comprises, in addition to the growth-enhancing molecule, at least one of:

collagen, fibronectin, and autologous plasma.

41. The method according to claim 39, comprising applying to the surface a layer that includes the growth-enhancing molecule and a separate layer that includes the at least one of:

collagen, fibronectin, and autologous plasma.

42. The method according to claim 1, wherein stimulating the CCP comprises:

during a low-serum time period, culturing the CCP in a culture medium comprising less than 10% serum; and

during a high-serum time period, culturing the CCP in a culture medium comprising greater than or equal to 10% serum.

48. The method according to claim 1, wherein stimulating the CCP comprises:

culturing the CCP in a first container during a first portion of a culturing period;

removing at least some cells of the CCP from the first container at the end of the first portion of the period; and

culturing, in a second container during a second portion of the period, the cells removed from the first container.

55. The method according to claim 1, wherein stimulating comprises culturing the CCP with at least one factor derived from a target tissue.

57. The method according to claim 1, wherein stimulating comprises co-culturing the CCP with a target tissue.

60. The method according to claim 57, wherein co-culturing comprises separating the target tissue from the CCP by a semi-permeable membrane.

61. The method according to claim 1, comprising deriving the CCP from peripheral blood.

62. The method according to claim 1, wherein stimulating the CCP comprises incubating the CCP in a container having a surface comprising an antibody.

63. The method according to claim 1, wherein stimulating the CCP comprises incubating the CCP in a container having a surface comprising autologous plasma.

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