Homozygous and heterozygous IDH1 gene-defective human astrocytoma cell lines

Patent metadata

Field	Value
Publication number	US-9695400-B2
Application number	US-201113878596-A
Country	US
Kind code	B2
Filing date	Oct 24, 2011
Priority date	Oct 22, 2010
Publication date	Jul 4, 2017
Grant date	Jul 4, 2017

Abstract

We provide IDH1 gene-defective cell lines (e.g., IDH1R132H heterozygous and IDH1R132H homozygous) derived from dissociated human astrocytoma samples. The cells can be used alone or in combination with each other or other cell types as a tool for determining the impact of IDH1R132H on cellular biology, tumorigenesis, and metabolic profiles. The cell lines may be used to test and identify therapeutic targets and to screen for molecular therapeutic agents.

First claim

We claim:

1. A set of isogenic, continuous human astrocytoma cell lines comprising at least two cell lines comprising:
(a) an astrocytoma cell whose genome comprises an IDH1 R132H allele and an IDH1 wild-type (WT) allele, and
(b) an astrocytoma cell whose genome comprises an IDH1 R132H allele and a disrupted IDH1 WT allele, wherein the disruption comprises an insertion of an exogenous synthetic cassette comprising either adeno-associated virus sequences or a neomycin resistance gene, wherein said cell lines grow indefinitely in culture as adherent cells.

2. A kit comprising in a divided container or multiple containers the set of cell lines of claim 1 .

3. An isolated and purified, continuous human astrocytoma cell line comprising astrocytoma cells, wherein the genome of the cells comprise an IDH1 R132H allele and a disrupted IDH1 WT allele, wherein the disruption comprises an insertion of a synthetic cassette, wherein said cells grow indefinitely in culture as adherent cells.

4. An isolated and purified, continuous human astrocytoma cell line comprising astrocytoma cells, wherein the genome of the cells comprise an IDH1 R132H allele and a disrupted IDH1 WT allele, wherein the disruption comprises an insertion of adeno-associated virus sequences, wherein said cells grow indefinitely in culture as adherent cells.

5. An isolated and purified, continuous human astrocytoma cell line comprising astrocytoma cells, wherein the genome of the cells comprise an IDH1 R132H allele and a disrupted IDH1 WT allele, wherein the disruption comprises an insertion of a neomycin resistance gene, wherein said cells grow indefinitely in culture as adherent cells.

6. The cell line of claim 3 made by a process comprising:
(a) enzymatically dissociating an astrocytoma tissue sample to produce dissociated cells,
(b) culturing the dissociated cells in stem cell medium,
(c) passaging the dissociated cells in stem cell medium five times,
(d) transferring the dissociated cells to a flask in a medium, whereby the cells adhere to the flask; and
(e) isolating the adherent cells in step (d), wherein the adherent cells consist of astrocytoma cells.

7. The cell line of claim 4 made by a process comprising:
(a) enzymatically dissociating an astrocytoma tissue sample to produce dissociated cells,
(b) culturing the dissociated cells in stem cell medium,
(c) passaging the dissociated cells in stem cell medium five times,
(d) transferring the dissociated cells to a flask in a medium, whereby the cells adhere to the flask; and
(e) isolating the adherent cells in step (d), wherein the adherent cells consist of astrocytoma cells.

8. The cell line of claim 5 made by a process comprising:
(a) enzymatically dissociating an astrocytoma tissue sample to produce dissociated cells,
(b) culturing the dissociated cells in stem cell medium,
(c) passaging the dissociated cells in stem cell medium five times,
(d) transferring the dissociated cells to a flask in a medium, whereby the cells adhere to the flask; and
(e) isolating the adherent cells in step (d), wherein the adherent cells consist of astrocytoma cells.

Assignees

• Yan Hai
• Bigner Darell
• Jin Genglin
• Univ Duke

Inventors

• Yan Hai
• Bigner Darell
• Jin Genglin

Classifications

• C12N5/0693 Primary

  Tumour cells; Cancer cells · CPC title