ESIBIe003-A-10

Not yet reviewed by Australian Stem Cell Registry

HES3 INS-GFP GCG-mCh, INSGFP/wGCGmCh/w hESC, MCRIe004-A-10

Details

Tissue & Disease as reported

Disease as reported (Genomic Modifications)

Genetic Information

Genotype Locus
GCG; INS
Modifications
Gene Knock-in Gene Knock-in

Associated Publications

Journal Year Article
Stem cell research 2021 An INSULIN-GFP/GLUCAGON-mCherry reporter line for the study of human pancreatic endocrine cell development

Line Custodianship

Cell Line Maintainer

Schiesser

Affiliated Institutions
  • Murdoch Children's Research Institute, Melbourne, Australia
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Cell Line Producer

Murdoch Children's Research Institute

Affiliated Institutions
  • Murdoch Children's Research Institute, Melbourne, Australia

Derivation Details

Ethics

Ethics Number: 33001A

Institution Human Research Ethics Council: Royal Children's Hospital Human Research Ethics Committee

Approval Date: To be verified

Modifications

Genomic Modifications

GFP was inserted into one allele of the INSULIN locus as described in Micallef et al 2012

Cytoband: cytoband location 11p15.5
Mutation Type: Gene Knock-in
Delivery Method: homologous_recombination

mCherry was inserted into one allele of the GLUCAGON locus as described in Labonne et al 2021

Cytoband: cytoband location 2q24.2
Mutation Type: Gene Knock-in
Delivery Method: talen

Quality Assurance

Genomic Characterisation

Passage Number: Not available

Karyotype: Normal

Karyotype Method: Molecular karyotyping by SNP array

Summary: Molecular karyotyping by SNP array (Illumina Infinium CoreExome-24; resolution 0.5 Mb). SNPduo comparison to parental hES3 INSULIN-GFP/w (single knock-in) line (>99.9% identity).

Microbiology and Virology Screening

Disease Result
Mycoplasma Negative

Characterisation of Undifferentiated Cells

Marker Method
POU5F1 (OCT-4) Immunostaining
CD9 Flow cytometry
EPCAM Flow cytometry
SSEA-4 Flow cytometry

Scorecard Results

Undifferentiated Cells

No available data for this cell line.

Pluripotency

No available data for this cell line.

Pluripotency Characterisation

Endoderm

In vitro directed differentiation

Assessed by: Reporter expression

Markers: INS;GCG

Mesoderm

In vitro directed differentiation

Assessed by: Flow cytometry

Markers: CD34;CD43

Ectoderm

In vitro directed differentiation

Assessed by: Immunostaining

Markers: TUBB3

Growth Characteristics

Culture Medium and Growth Conditions

CO2 concentration: Unavailable

O2 concentration: Unavailable

Medium Items:

Passage Method: Enzymatically