MCRIi010-A-1

Flagged for further review

Alternative Names: ACTA1tdTom; PB010.5-ACTA1tdTom

Details

Description

This induced pluripotent stem cell line (MCRIi010-A-1) is a subclone of MCRIi010-A, which was originally reprogrammed from male peripheral blood mononuclear cells using non-integrating sendai viral vectors. MCRIi010-A-1 was generated by homozygous insertion of the tdTomato fluorescent protein and a P2A peptide at the C-terminal of ACTA1. MCRIi010-A-1 is described as having a normal karyotype. Expression of OCT4, EPCAM, TRA-1-81 and SSEA-4 in undifferentiated cells, and directed differentiation to endoderm, mesoderm, ectoderm and skeletal muscle myotubes have been reported.

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

Line Custodianship

Cell Line Maintainer

Peter Houweling

Affiliated Institutions
  • Murdoch Children's Research Institute, Melbourne, Australia
View all cell lines from this group

Cell Line Producer

Murdoch Children's Research Institute

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

Source

Parental Cell Line

MCRIi010-A

Donor Derived

Age: 20-24

Biological Sex: Male

Derivation Details

Induced Pluripotent Cell Derivation Details

Source Cell Type: EFO:0004905
Label: induced pluripotent stem cell
Definition: Induced pluripotent stem cells (iPS cells or iPSCs) are a type of pluripotent stem cell artificially derived from a non-pluripotent cell. Various methods exist to revert cells to pluripotency such as reprogramming mediated through a mature metaphase II oocyte as in somatic cell nuclear transfer.

Source Cell Origin: N/A

Derivation Year:

Ethics

Ethics Number: 35121A

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

Approval Date: None

Modifications

Genomic Modifications

ACTA1

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

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

Microbiology and Virology Screening

Disease Result
Mycoplasma Negative

Characterisation of Undifferentiated Cells

Marker Method
POU5F1 (OCT-4) ddPCR
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: In vitro directed differentiation

Markers: SOX17

Mesoderm

In vitro directed differentiation

Assessed by: In vitro directed differentiation

Markers: TBXT, CXCR4

Ectoderm

In vitro directed differentiation

Assessed by: In vitro directed differentiation

Markers: NES, PAX6

Growth Characteristics

Culture Medium and Growth Conditions

CO2 concentration: 5%

O2 concentration: Unavailable

Medium Items:

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

Passage Method: Enzyme-free cell dissociation