AST22-C, AST23-C

General

Cell Line

hPSCreg Name
EDi001-A-1
Alternative name(s)
AST22-C, AST23-C
Cell line type Human induced pluripotent stem cell (hiPSC)
Last update 14th May 2022
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Provider

Generator University of Edinburgh (ED)
Owner College of Medicine and Veterinary Medicine
Distributors
Derivation country United States

External Databases

BioSamples SAMEA3323836
Cellosaurus CVCL_LE51
ECACC 66540166
EBiSC EDi001-A-1
CLO CLO_0102712
Wikidata Q54831972

General Information

Publications
Projects
* Is the cell line readily obtainable for third parties?
Yes
Research use: allowed
Clinical use: allowed
Commercial use: allowed
Subclone of

Donor Information

General Donor Information

Sex female

Phenotype and Disease related information (Donor)

Diseases A disease was diagnosed.
Parkinson disease
This is a PD line, with the control being EDi002-A lines and CRISPR/Cas9-corrected EDi001-A-1, EDi001-A-2, EDi001-A-3 and EDi001-A-4
The donor is a carrier of a disease-associated mutation and affected.
Genetic variants
SNCA (target)
4q22.1
Heterozygous
The donor carries a triplication of the alpha-synuclein gene, resulting in 4 copies of SNCA. The copies of SNCA are situated in a heterozygous triplication configuration. See Figure 1 of Petrucci, 2015 for a graphic representation of the heterozygous triplication.
Disease associated phenotypes
  • Severe PD with dementia
Family history Strong family history of Parkinson’s disease due to autosomal dominant inheritance of SNCA triplication
Is the medical history available upon request? Y Mov Disord. 2011 Sep;26(11):2134-6. doi: 10.1002/mds.23776

Karyotyping (Donor)

Has the donor karyotype been analysed?
No

Donor Relations

Other cell lines of this donor
All cell lines of this donor's relatives
Has daughter:

External Databases (Donor)

BioSamples SAMEA3319991

Ethics

Also have a look at the ethics information for the parental line EDi001-A .
For generation of the cell line, who was the supplier of any recombined DNA vectors or commercial kits used?

hIPSC Derivation

General

The source cell information can be found in the parental cell line EDi001-A.

Reprogramming method

Vector type Integrating
Vector Virus (Retrovirus)
Genes
Is the used vector excisable?
No
Absence of reprogramming vector(s)?
Unknown
Reprogramming vectors silenced?
Yes
Methods used
Immunostaining, RT-PCR

Vector free reprogramming

Other

Derived under xeno-free conditions
No
Derived under GMP?
No
Available as clinical grade?
No

Culture Conditions

Surface coating Laminin
Feeder cells
No
Passage method Enzymatically
Accutase
O2 Concentration 95 %
CO2 Concentration 5 %
Medium Essential 8™
Supplements
Rock inhibitor added while passaging 10 nM

Characterisation

Analysis of Undifferentiated Cells
Marker Expressed Immunostaining RT-PCR FACS Enzymatic Assay Expression Profiles
TRA 1-60
Yes
SSEA-4
Yes
POU5F1 (OCT-4)
Yes
SSEA-1
No
Differentiation Potency
Endoderm
Ont Id: UBERON_0000925
In vitro spontaneous differentiation
Mesoderm
Ont Id: UBERON_0000926
In vitro spontaneous differentiation
Ectoderm
Ont Id: UBERON_0000924
In vitro spontaneous differentiation

Microbiology / Virus Screening

HIV 1 Negative
HIV 2 Negative
Hepatitis B Negative
Hepatitis C Negative
Mycoplasma Negative

Certificate of Analysis

Is there a certificate of analysis available?
Yes
Passage: 58

Genotyping

Karyotyping (Cell Line)

Has the cell line karyotype been analysed?
No

Other Genotyping (Cell Line)

Genetic Modification

Disease/phenotype related modifications
Parkinson disease
Genetic modifications
SNCA (target)
Isogenic modification
4q22.1
CRISPR/Cas was employed to create a mutation in Exon2, disrupting the ATG start site of SNCA. The site targeted for editing was sequenced for 8 clones; however, in these 8 clones, none of the sites had been modified. Therefore, it appears that the cell line had not been edited. The clone was not further examined for off-target modifications.
This clone was picked in parallel to other Nickase pair targeted clones (EDi001-A2/A3 and A4) but no SNCA alleles were lost in this clone. Therefore all 4 alleles remain, and it retains the donor heterozygous triplication of SNCA.