ΔINK4 T2D risk region hiPSC

General#

Cell Line

hPSCreg Name HMGUi001-A-5
Alternative name(s)
ΔINK4 T2D risk region hiPSC
Cell line type Human induced pluripotent stem cell (hiPSC)
Last update 30th June 2020
User feedback
No feedback available yet.

Login to share your feedback, experiences or results with the research community.

Provider

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

External Databases

BioSamples SAMEA7178021
CLO CLO_0102765

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.
Disease associated phenotypes no phenotypes
Family history no
Is the medical history available upon request? no
Is clinical information available? no

Karyotyping (Donor)

Has the donor karyotype been analysed?
No

Other Genotyping (Donor)

Is there genome-wide genotyping or functional data available?
No

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? No
For generation of the cell line, who was the supplier of any recombined DNA vectors or commercial kits used? The targeting vector was generated at IDR, and contained the PU6-(BbsI) sgRNA_CAG-Cas9-GFP-bpA plasmid, Addgene ID86985.
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 an Ethics Committee is oblgatory.

hIPSC Derivation#

General

The source cell information can be found in the parental cell line HMGUi001-A.
Passage number reprogrammed 32

Reprogramming method

Vector type Non-integrating
Vector HMGUi001-A-1 was targeted with PU6-(BbsI) sgRNA_CAG-Cas9-GFP-bpA plasmid, Addgene ID8698,5 to make a large deletion at the INK4 locus. This vector expresses Cas9 enzyme and sgRNAs.

Vector free reprogramming

Other

Selection criteria for clones FACS sorting of GFP expressing cells, genomic DNA exctraction from GFP+ single cell clones, screening for the 8kb deletion by conventional PCR with FA-RB and FC-RC primer pairs, identification of wild type and biallelic clones for the large deletion by a 1910 bp wild type-specific PCR product and a 750 bp deletion-specific PCR product, confirmation of the precise deletion of the target region by Sanger sequencing of the PCR products
Derived under xeno-free conditions
No
Derived under GMP?
No
Available as clinical grade?
No

Culture Conditions#

Surface coating Matrigel/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-fee
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?
No
Has Rock inhibitor (Y27632) been used at thaw previously with this cell line?
No

Characterisation#

Analysis of Undifferentiated Cells
Marker Expressed Immunostaining RT-PCR FACS Enzymatic Assay Expression Profiles
SOX2
Yes
POU5F1 (OCT-4)
Yes
StemMACS Trilineage Differentiation plus immunostaining
Differentiation Potency
Endoderm
Ont Id: UBERON_0000925
Mesoderm
Ont Id: UBERON_0000926
Ectoderm
Ont Id: UBERON_0000924

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
INK4 (target)
Variant
9P21.3
Deletion of the INK4-T2D risk region at both alleles (homozygous or biallelic)