Identification of cells for transplantation

Details Identification of cells for transplantation Identification of cells for transplantation

2000-10-25

2002-10-15

Fredman, Jeffrey (Department: 1656)

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Recombinant dna technique included in method of making a...

C435S006120, C435S091200

Reexamination Certificate

active

06465215

ABSTRACT:

FIELD OF THE INVENTION
This invention relates to the identification of cells suitable for transplantation into a vertebrate brain. More particularly, this invention relates to the identification of multipotent neural cells which are able to repair neural damage following transplantation into the brain.
BACKGROUND TO THE INVENTION
There is an increasing awareness that damage to a vertebrate brain can be repaired using cell transplantation technology. Typically, the cell transplanted into a damaged brain will be a neural stem cell, e.g. a pluripotent neuroepithelial stem cell which is capable of differentiating into a cell with a neural cell phenotype.
For example, Sinden et al., Neuroscience (1997) 81:599-608 , discloses that conditionally-immortalised hippocampus neuroepithelial stem cells can be used to improve spatial learning after transplantation into the ischaernia-leisoned hippocampus. See also WO-A-97/10329.
However, it has been found that not all neural stem cells can be transplanted for successful repair of neural damage.
For example, while MHP36 cells (Sinden et al., supra) do aid repair, an apparently similar cell line, MHP15, fails to repair. This difference emerges despite the fact that both MHP cell lines were generated from the same tissue source (namely hippocampus), and both are multipotent neural precursor cell lines, i.e. both cell lines have the capacity to generate a full complement of brain cell types, including neurons, astrocytes and oligodendrocytes.
Therefore, it would be beneficial if it were possible to identify at an early state, the cells that were suitable for transplantation, or to have the ability to modify cell lines in order to achieve successful transplantation and repair.
SUMMARY OF THE INVENTION
The present invention is based on the realisation that cells that are suitable for transplantation and repair may be identified on the basis of their gene expression profile.
The present invention employs, for example, a technique termed differential display (DD) to investigate the differences between repairing and non-repairing cell lines. Differential display is used to visualise the differences in gene expression between two or more cell lines, and it has been found that cell lines that repair have very similar profiles of gene expression but very different profiles from those cells that do not repair. This represents a major, unexpected discovery because, apart from their capacity to repair, the cell lines are usually remarkably similar, in morphology, growth characteristics and growth factor responsivity.
According to one aspect of the present invention, a method for selecting a cell suitable for transplantation into a damaged vertebrate brain comprises:
(i) isolating cells that are, or are capable of differentiating into, a cell with a neural cell phenotype;
(ii) obtaining the gene expression profile of the cells;
(iii) comparing the expression profile of the cells with that from a control cell known to be suitable for transplantation; and
(iv) selecting those cells with a similar expression profile to that of the control.
In one embodiment of the invention, the control cell is from the MHP 36 cell line. Step (ii) may be carried out by differential display.
The present invention is not only useful for identifying suitable cells for transplantation, but may be used to identify genes or gene products that may be involved in determining whether or not a neural cell can repair, or not.
According to a further aspect of the present invention, a method for identifying a gene involved in determining whether or not a neural cell can aid repair on transplantation into a damaged brain, comprises:
(i) isolating cells that are, or are capable of differentiating into, a cell with a neural cell phenotype;
(ii) obtaining the gene expression profile of the cells;
(iii) comparing the expression profile of the cells with that from a control cell line known to be suitable for transplantation; and
(iv) isolating those genes that are the same (or different) from those expressed by the control.
Using this method, it has been possible to identify a number of genes that are expressed by repairing cell lines but not non-repairing cell lines (and vice versa).
According to a third aspect, a method for selecting a cell suitable for transplantation into a damaged brain, comprises determining the presence of any of the gene sequences identified herein as SEQ ID NOS. 1 to 5.
BRIEF DESCRIPTION OF THE SEQUENCES
SEQ ID NOS. 1-7 are polynucleotide sequences useful according to the subject invention.


REFERENCES:
patent: 5753491 (1998-05-01), Major et al.
patent: 5851832 (1998-12-01), Weiss et al.
patent: 5958767 (1999-09-01), Snyder et al.
patent: 8903872 (1989-05-01), None
patent: 9109936 (1991-07-01), None
patent: 9301275 (1993-01-01), None
patent: 9615226 (1996-05-01), None
patent: 9710329 (1996-09-01), None
patent: WO-97/10329 (1997-03-01), None
patent: 9829565 (1997-12-01), None
patent: 9915700 (1998-09-01), None
patent: 9927076 (1998-11-01), None
patent: 9946385 (1999-03-01), None
Utans U et al. Chronic cardiac rejection: Identification of five upregulated genes in transplanted hearts by differential mRNA display. Proc.Natl.Acad.Sci.USA., 91: 6463-6467, 1994.*
Gotz M et al. Pax6 controls radial glia differentiation in the cerebral cortex. Neuron, 21:1031-1044, 1998.*
Marra M et al. vi78b11.x1 stratagene mouse testis (3937308) Mus musculus cDNA clone. Accession No. AI507135. EST database search report, Mar. 11, 1999.*
U.S. application No. 09/663,537, Jat, filed Sep. 15, 2000.
U.S. application No. 09/537,617, Hodges, filed Mar. 29, 2000.
U.S. application No. 09/672,606, Sinden et al., filed Sep. 28, 2000.
U.S. application No. 09/760,274, Sinden et al., filed Jan. 12, 2001.
Snyder, Evan Y., David L. Deitcher, Christopher Walsh, Susan Arnold-Aldea, Erika A. Hartwieg, Constance L. Cepko (Jan. 10, 1992) “Multipotent Neural Cell Lines Can Engraft and Particpate in Development of Mouse Cerebellum”Cell68:33-51.
Snyder, Evan Y. and John H. Wolfe (1996) “Central nervous system cell transplantation: a novel therapy for storage diseases?”Curr. Opin. Neurol.9:126-136.
Snyder, Evan Y., Cliff Yoon, Jonathan D. Flax, Jeffrey D. Macklis (Oct. 14, 1997) “Multipotent neural precursors can differentiate toward replacement of neurons undergoing targeted apoptotic degeneration in adult mouse neocortex”Proc. Natl. Acad. Sci.94(21):11663-11668.
Utans, Ulrike, Peng Liang, Lauri R. Wyner et al. (Jul. 1994) “Chronic cardiac rejection: Identification of five upregulated genes in transplanted hearts by differential mRNA display”Proc. Natl. Acad. Sci. USA91:6463-6467.
Götz, Magdalena, Anastassia Stoykova, Peter Gruss (Nov. 1998) “Pax6 Controls Radial Glia Differentiation in the Cerebral Cortex”Neuron21:1031-1044.
Sinden, J.D., F. Rashid-Doubell, T.R. Kershaw, et al. (1997) “Recovery of Spatial Learning By Grafts Of A Conditionally Immortalized Hippocampal Neuroepithelial Cell Line Into The Ischaemia-Lesioned Hippocampus”Neurosciencevol. 81 (3):599-608.
Fernandez, Anibal Smith, Claude Pieau, Jacques Repérant et al. (1998) “Expression of the Emx-1 and Dlx-1 homeobox genes define three molecularly distinct domains in the telencephalon of mouse, chick, turtle and frog embryos: implications for the evolution of telencephalic subdivisions in amniotes”Development125:2099-2111.
Puelles, Luis and John L.R. Rubenstein (1993) “Expression patterns of homeobox and other putative regulatory genes in the embryonic mouse forebrain suggest a neuromeric organization”TINS16(11):472-479.
McKay et al. (1990) “Mechanisms regulating Cell Number and Type in the Mammalian Central Nervous System” Cold Spring Harbor Symposia on Quantitative Biology, vol. 55, pp. 291-301.
McKay et al. (1993) “Immortalized Stem Cells Form the Central Nervous System” C.R. Acad. Sci. Pars, Sciences De La Vie, vol. 316, pp. 1452-1457.
Okabe et al. (1996) “Development of Neuronal Precursor Cells and Functional Postmitotic Neurons from Embryonic Stem Cells In Vitro” Mechanisms of Development, vol. 59, pp. 89-102.
Sanberg et al. (Feb., 1998) Pro

Affiliated with

Also associated with

Rating

Identification of cells for transplantation does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Identification of cells for transplantation, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Identification of cells for transplantation will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-2992597