MCRIi024-A-1

Not yet reviewed by Australian Stem Cell Registry

ACTA1.Bbc

Details

Tissue & Disease as reported

Disease as reported (Genomic Modifications)

Genetic Information

Genotype Locus
ACTA1
Polymorphism
Homozygous: unspecified_reference_ACTA1:c.443=(p.Gly148=)
Modifications
Isogenic modification

Associated Publications

Journal Year Article
Stem Cell Research 2021 Generating an iPSC line (with isogenic control) from the PBMCs of an ACTA1 (p.Gly148Asp) nemaline myopathy patient.
Stem cell research 2021 Generating an iPSC line (with isogenic control) from the PBMCs of an ACTA1 (p.Gly148Asp) nemaline myopathy patient

Line Custodianship

Cell Line Maintainer

North

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

Derivation Details

Induced Pluripotent Cell Derivation Details

Source Cell Type: N/A

Source Cell Origin: N/A

Derivation Year:

Ethics

Ethics Number: 38192

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

Approval Date: To be verified

Modifications

Genomic Modifications

Homozygous: unspecified_reference_ACTA1:c.443=(p.Gly148=)

Correction of heterozygous ACTA1 c.443G>A (p.Gly148Asp) variant present in the parental line (MCRIi024-A). A synonymous change was also introduced (ACTA1 p.Ser147Ser). Targeted corrected clones were identified by allele-specific PCR and changes were confirmed by Sanger Sequencing.

Cytoband: 1q42.13
Mutation Type: Isogenic modification
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 array ((Illumina Infinium GSA-24 v1.0; resolution 0.5 Mb). SNPduo comparison to parental cell (>99.9% identity).

Microbiology and Virology Screening

Disease Result
Mycoplasma Negative

Characterisation of Undifferentiated Cells

Marker Method
OCT3/4 Flow cytometry
SSEA-4 Flow cytometry
TRA-1-60 Flow cytometry
NANOG Immunostaining
POU5F1 (OCT-4) Immunostaining
POU5F1 (OCT-4) ddPCR

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: ddPCR

Markers: SOX17

Mesoderm

In vitro directed differentiation

Assessed by: ddPCR

Markers: FOXC2;MEOX1

Ectoderm

In vitro directed differentiation

Assessed by: ddPCR

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