MCRIi010-A-1

Extracted from hPSCreg

ACTA1tdTom, PB010.5-ACTA1tdTom

Details

Tissue & Disease as reported

Disease as reported (Genomic Modifications)

Genetic Information

Genotype Locus
ACTA1
Polymorphism
tm(ACTA1t/t:ACTA1-P2A-tdTomato)
Modifications
Gene Knock-in

Associated Publications

Journal Year Article
Stem cell research 2024 Generation of a human ACTA1-tdTomato reporter iPSC line using CRISPR/Cas9 editing
Stem Cell Research 2024 Generation of a human ACTA1-tdTomato reporter iPSC line using CRISPR/Cas9 editing.

Line Custodianship

Cell Line Maintainer

Houweling-MCRI

View all cell lines from this group

Cell Line Producer

Murdoch Children's Research Institute

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

Derivation Details

Induced Pluripotent Cell Derivation Details

Source Cell Type: CL:0000765
Label: erythroblast
Definition: A nucleated precursor of an erythrocyte that lacks hematopoietic lineage markers.

Source Cell Origin: UBERON:0000178
Label: blood
Definition: A fluid that is composed of blood plasma and erythrocytes.

Derivation Year:

Ethics

Ethics Number: Holding Entry-MCRIi010-A

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

Approval Date: To be verified

Modifications

Genomic Modifications

tm(ACTA1t/t:ACTA1-P2A-tdTomato)

Homozygous insertion of sequences encoding the tdTomato fluorescent protein and a P2A peptide at the ACTA1 locus.

Cytoband: 1q42.13
Mutation Type: Gene Knock-in
Delivery Method: CRISPR/Cas9

Quality Assurance

Genomic Characterisation

Passage Number: Not available

Karyotype: arr(1-22)x2,(XY)x1

Karyotype Method: Molecular karyotyping by SNP array

Summary: Molecular karyotyping by SNP arrays (Illumina Infinium GSA-24 v3.0 (resolution 0.5Mb) and Infinium Global Diversity Array with Cytogenetics-8 (GDACyto-8) v1.0 (resolution 0.2 Mb)). SNPduo comparison to parental cell (>99.9% identity).

Microbiology and Virology Screening

Disease Result
Mycoplasma Negative

Characterisation of Undifferentiated Cells

Marker Method
OCT4 ddPCR
CD326(EPCAM) Flow cytometry
SSEA-4 Flow cytometry
TRA-1-81 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: Immunostaining

Markers: SOX17

Mesoderm

In vitro directed differentiation

Assessed by: ddPCR

Markers: Brachyury; CXCR4

Ectoderm

In vitro directed differentiation

Assessed by: Immunostaining

Markers: NES; PAX6

Growth Characteristics

Culture Medium and Growth Conditions

CO2 concentration: 5%

O2 concentration: Unavailable

Medium Items:

  • Base Coat: Matrigel - Corning
  • Base Medium: Essential 8 Medium - Thermo Fisher Scientific Inc.

Passage Method: Enzyme-free cell dissociation