STBCi004-B-1

SFC832-03-06 LRRK2WT/WT C47

Gene-edited iPSC line

At European Collection of Authenticated Cell Cultures (ECACC)
Timepoint: confluency
Magnification: 4x
Timepoint: confluency
Magnification: 10x
A CLIP contains information about a cell line including any specific third party obligations relating to, for example, licensing obligations or the donor consent which affect the use of the cell line.
A batch specific Certificate of Analysis will be available to download once you receive your EBiSC iPSC line.

General#

Cell Line

hPSCreg Name STBCi004-B-1
Alternative name(s)
SFC832-03-06 LRRK2WT/WT C47
Cell line type Human induced pluripotent stem cell (hiPSC)
Similar lines

Provider

Depositor StemBANCC (STBC)
Distributors
EBiSC
European Collection of Authenticated Cell Cultures (ECACC)

External Databases

hPSCreg STBCi004-B-1
BioSamples SAMEA5859478
ECACC 66541204
CLO CLO_0102950
Cellosaurus CVCL_A8X1
Wikidata Q102114943

General Information

Publications View all related publications on hPSCreg (1)
* Is the cell line readily obtainable for third parties?
Yes
Research use: allowed
Clinical use: not allowed
Commercial use: not allowed
Subclone of

Donor Information#

General Donor Information

Sex female

Phenotype and Disease related information (Donor)

Diseases A disease was diagnosed.
Parkinson disease (primary disease)
The donor is a carrier of a disease-associated mutation and affected.
Genetic variants
LRRK2 (target)
12q12
NM_198578.3:c.6055G>A
NP_940980.3:p.Gly2019Ser
LRRK2 G2019S

Donor Relations

Other cell lines of this donor

External Databases (Donor)

BioSamples SAMEA104129730

hIPSC Derivation#

General

The source cell information can be found in the parental cell line STBCi004-B.

Reprogramming method

Vector type Non-integrating
Vector Sendai virus
Genes
Is reprogramming vector detectable?
No
Methods used
PCR

Vector free reprogramming

Other

Derived under xeno-free conditions
Unknown
Derived under GMP?
Unknown
Available as clinical grade?
Unknown

Culture Conditions#

Latest released batch

Culture medium mTeSR1
Passage method EDTA
Surface coating Matrigel
O2 concentration 21
CO2 concentration 5
Temperature 37 °C
The following are the depositor culture conditions, they do not refer to any specific batch.
Surface coating Matrigel/Geltrex
Feeder cells
No
Passage method Enzyme-free cell dissociation
EDTA
Medium mTeSR™ 1

Characterisation#

Analysis of Undifferentiated Cells
Marker Expressed Immunostaining RT-PCR FACS Enzymatic Assay Expression Profiles
NANOG
Yes
TRA 1-60
Yes
POU5F1 (OCT-4)
Yes
SSEA-4
Yes
Spontaneous EB differentiation and qPCR for trilineage markers
Differentiation Potency
Endoderm
Ont Id: UBERON_0000925
In vitro spontaneous differentiation
Marker Expressed
CXCR4
Yes
GATA6
Yes
SOX17
Yes
Mesoderm
Ont Id: UBERON_0000926
In vitro spontaneous differentiation
Marker Expressed
VIM
Yes
DCN
Yes
MLX1
Yes
Ectoderm
Ont Id: UBERON_0000924
In vitro spontaneous differentiation
Marker Expressed
HES5
Yes
NEUROD1
Yes
PAX6
Yes

Microbiology / Virus Screening

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

Sterility

Inoculation for microbiological growth No Contaminants Detected
Mycoplasma Not Detected
Viability Viable post-cryopreservation

Genotyping#

Karyotyping (Cell Line)

Has the cell line karyotype been analysed?
Yes
Karyotype abnormalities: none compared to fibroblasts
Passage number: 40
Karyotyping method: Molecular karyotyping by SNP array
http://

Other Genotyping (Cell Line)

Genetic Modification#

Disease/phenotype related modifications
Parkinson disease
Genetic modifications
LRRK2 (target)
Isogenic modification
12q12
The G2019S variant (NM_198578.3:c.6055G>A) in the gene LRRK2 was normalized by gene editing, resulting in the wt form: NM_198578.3:c.6055G. CRISPR/Cas9-mediated double-strand break generated close to G2019S mutation and homology-directed repair with donor sequence. Homology arms generated by amplification of LRRK2 sequence in mutant allele to maintain isogeneic sequence. The G2019S mutation was repaired (G>A), and other silent mutations introduced including mutated protospacer adjacent motifs to prevent recutting of repaired sequence, and a PstI site for identification of repaired clones. See Certificate of Analysis.
Repaired