HMGUi001-A-65

CCKBRΔ12/KO

The cell line is not validated yet.

General

Cell Line

hPSCreg name HMGUi001-A-65
Cite as:
HMGUi001-A-65
Alternative name(s)
CCKBRΔ12/KO
Cell line type Human induced pluripotent stem cell (hiPSC)
Similar lines No similar lines found.
Last update 2nd April 2026
Notes For the use of the cell line a material transfer agreement with HMGU is obligatory and can be requested from material-transfer@helmholtz-munich.de.
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Provider

Generator Institute of Diabetes and Regeneration Research (IDR)
Owner Institute of Diabetes and Regeneration Research (IDR)
Distributors
Derivation country Germany

External Databases

BioSamples SAMEA122093052

General Information

* Is the cell line readily obtainable for third parties?
No
Subclone of

Donor Information

General Donor Information

Sex female
Ethnicity Caucasian

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

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 SAMEA5696688

Ethics

Also have a look at the ethics information for the parental line HMGUi001-A .
Is there an MTA available for the cell line? Yes
For generation of the cell line, who was the supplier of any recombined DNA vectors or commercial kits used? Vectors were generated at IDR, pU6-(BbsI)-sgRNA-CAG-Cas9-Venus-bpA plasmid (Addgene plasmid #86986) was used.
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? Yes
Constraints for use or distribution According to donor consent the line must not be used for commercial purposes. For research purposes project-specific review by a medical Ethics Committee is obligatory.

hIPSC Derivation

General

The source cell information can be found in the parental cell line HMGUi001-A.
Passage number reprogrammed 26

Reprogramming method

Vector type Non-integrating
Vector The CCKBR locus was targeted by homologous recombination and CRISPR/Cas9 technology using histone 2B (H2B)-Venus-3xHA-tag targeting vector. We cloned the sequence of the (H2B)-Venus-3xHA-tag into a targeting vector and used the 897 bp upstream and 833 bp downstream of the CCKBR start and stop codon respectively as 5′ and 3′ homology arms (HA). A pair of gRNAs introducing dsDNA breaks 8 bp downstream of the start codon and 2 bp upstream of the stop codon of CCKBR were cloned into the pu6-sgRNA-CAG-Cas9-VenusbpA expression vector.
Is reprogramming vector detectable?
Yes

Vector free reprogramming

Other

Selection criteria for clones Transfected cells expressed Venus and were sorted by FACS. The genotype of the clones was screened by PCR and verified by Sanger sequencing.
Derived under xeno-free conditions
Yes
Derived under GMP?
No
Available as clinical grade?
No

Culture Conditions

Surface coating Geltrex
Feeder cells
No
Passage method EDTA
O2 Concentration 21 %
CO2 Concentration 5 %
Medium Other medium:
Base medium: StemMACS iPS-Brew XF medium
Main protein source: Xeno-free
Serum concentration: 0 %
Has Rock inhibitor (Y27632) been used at passage previously with this cell line?
Yes
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 Expressed Immunostaining RT-PCR Flow Cytometry Enzymatic Assay Expression Profiles
POU5F1 (OCT-4)
Yes
SSEA-4
Yes
TRA 1-60
Yes
Oct-3
Yes
Differentiation Potency
Endoderm
Ont Id: UBERON_0000925
Marker Expressed
SOX17
Yes
FOXA2
Yes
Mesoderm
Ont Id: UBERON_0000926
Marker Expressed
SM22-alpha
Yes
Ectoderm
Ont Id: UBERON_0000924
Marker Expressed
Nestin
Yes

Microbiology / Virus Screening

Mycoplasma Negative

Genotyping

Karyotyping (Cell Line)

Has the cell line karyotype been analysed?
Yes
46,XX
Karyotyping method: G-Banding

Other Genotyping (Cell Line)

Genetic Modification

Genetic modifications not related to a disease
CCKBR (target)
Variant
Chromosome 11, CCKBR gene, exon 5
Heterozygous
The CCKBRKO H2B venus KI hiPSC line presented here was generated by hetrozygous replacement of CCKBR exon1-5 by H2B-Venus sequence resulting in deletion of 12 bp after ATG in CCKBR in one allele and expression of H2B-Venus under CCKBR promoter activity in the other allele.