WAe009-A-O

General

Cell Line
hPSCreg Name	WAe009-A-O
Alternative name(s)	CRX-GFP WAe009
Cell line type	Human embryonic stem cell (hESC)
Last update	27th April 2023
User feedback	Written by on No feedback available yet. Login to share your feedback, experiences or results with the research community.
Provider
Generator	University of Newcastle (UNEW)
Owner	University of Newcastle (UNEW)
Distributors	University of Newcastle (UNEW)
Derivation country	United Kingdom
External Databases
BioSamples	SAMEA112970166
CLO	CLO_0104738
General Information
Projects	HYPOIMMUNE: Off-the-shelf hypoimmunogenic photoreceptors for treatment of blinding retinal disease
* Is the cell line readily obtainable for third parties?	Yes Research use: allowed Clinical use: not allowed Commercial use: allowed
Subclone of	WAe009-A

Donor Information

General Donor Information

Sex

female

Ethnicity

N/A

Phenotype and Disease related information (Donor)

Diseases

No disease was diagnosed.

Family history

NO

Is the medical history available upon request?

NO

Is clinical information available?

NO

Karyotyping (Donor)

Has the donor karyotype been analysed?

Unknown

Other Genotyping (Donor)

Is there genome-wide genotyping or functional data available?

No

Donor Relations

Other cell lines of this donor

External Databases (Donor)

BioSamples

SAMEA7768918

Ethics

Also have a look at the ethics information for the parental line WAe009-A .

Is there an MTA available for the cell line?	No
Are you aware of any constraints on the use or distribution of the cell line from the owner or any parties identified in the query above?	No

hESC Derivation

The source cell information can be found in the parental cell line WAe009-A.

Culture Conditions

Surface coating	Matrigel/Geltrex
Feeder cells	No
Passage method	Enzymatically Versene
O2 Concentration	21 %
CO2 Concentration	5 %
Medium	mTeSR™ 1 PSC_culture_with_mTesr1_media.pdf
Has Rock inhibitor (Y27632) been used at passage previously with this cell line?	No
Has Rock inhibitor (Y27632) been used at cryo previously with this cell line?	Yes
Has Rock inhibitor (Y27632) been used at thaw previously with this cell line?	Yes

Characterisation

Analysis of Undifferentiated Cells

Marker Expression

Marker	Expressed	Immunostaining	RT-PCR	FACS	Enzymatic Assay	Expression Profiles
SSEA-4	Yes	–

Differentiation Potency

Endoderm

Mesoderm

Ectoderm

Genotyping

Karyotyping (Cell Line)
Has the cell line karyotype been analysed?	Yes 46 XX Collin_et_al_Suppl__Figure_3_copy.tif Passage number: 50 Karyotyping method: G-Banding
Other Genotyping (Cell Line)

Genetic Modification

Genetic modifications not related to a disease

Type of modification

Gene knock-in

Chromosome location

19q13.33

Chromosome location (transgene)

19q13.33

Free text

ZFNs designed to cleave in the 3’ UTR of CRX were transfected into hESCs along with a donor construct containing homology to the target region, eGFP reporter and a puromycin selection cassette. Following selection, PCR and sequencing analysis of antibiotic resistant clones indicated targeted integration of the reporter cassette at the 3’ of the CRX gene, generating a CRX-GFP fusion. Further analysis of a clone exhibiting homozygote integration of the GFP reporter was conducted suggesting genomic stability was preserved and no other copies of the targeting cassette were inserted elsewhere within the genome.

Delivery method

Homologous Recombination

CRX-GFP WAe009