MCRIi001-A-2

Not yet reviewed by Australian Stem Cell Registry

PB001-SOX9tdTom, MCRIi001-A-SOX9tdTom

Details

Tissue & Disease as reported

Disease as reported (Genomic Modifications)

Genetic Information

Genotype Locus
SOX9
Polymorphism
tm(SOX9t/+:SOX9-T2A-tdTomato)
Modifications
Transgene Expression 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
Proceedings of the National Academy of Sciences of the United States of America 2023 Modeling human skeletal development using human pluripotent stem cells
Seminars in cell & developmental biology 2022 Developmental principles informing human pluripotent stem cell differentiation to cartilage and bone
Stem cell research 2020 Generation of a SOX9-tdTomato reporter human iPSC line, MCRIi001-A-2, using CRISPR/Cas9 editing
Computational and structural biotechnology journal 2020 Probing pluripotency gene regulatory networks with quantitative live cell imaging
Stem cell research 2020 CRISPR/Cas9 gene editing of a SOX9 reporter human iPSC line to produce two TRPV4 patient heterozygous missense mutant iPSC lines, MCRIi001-A-3 (TRPV4 p.F273L) and MCRIi001-A-4 (TRPV4 p.P799L)
Stem cell research 2020 Generation of a heterozygous COL2A1 (p.R989C) spondyloepiphyseal dysplasia congenita mutation iPSC line, MCRIi001-B, using CRISPR/Cas9 gene editing

Line Custodianship

Cell Line Maintainer

Lamandé

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: 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: 35121A

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

Approval Date: To be verified

Modifications

Genomic Modifications

Td-Tomato fluorescence reporter gene insertion downstream of SOX9 gene

Mutation Type: Transgene Expression
Delivery Method: crispr

tm(SOX9t/+:SOX9-T2A-tdTomato)

Heterozygous insertion of a tdTomato reporter gene with upstream T2A sequence at the stop codon of SOX9 exon 3.

Cytoband: 17q24.3
Mutation Type: Gene Knock-in
Delivery Method: CRISPR/Cas9

Quality Assurance

Genomic Characterisation

Passage Number: 2

Karyotype: Molcular karyotype arr(1-22)x2(XY)x1

Karyotype Method: Molecular karyotyping by SNP array

Microbiology and Virology Screening

Disease Result
Mycoplasma Negative

Characterisation of Undifferentiated Cells

Marker Method
NANOG Immunostaining
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 spontaneous differentiation

Assessed by: Immunostaining

Markers: HNF4A

Mesoderm

In vitro spontaneous differentiation

Assessed by: Immunostaining

Markers: COL2A1

Ectoderm

In vitro spontaneous differentiation

Assessed by: Immunostaining

Markers: MAP2

Growth Characteristics

Culture Medium and Growth Conditions

CO2 concentration: 5%

O2 concentration: 95%

Medium Items:

  • Base Medium: KnockOut DMEM/F12 - Thermo Fisher Scientific Inc.
  • Serum: KnockOut Serum Replacement - Thermo Fisher Scientific Inc.
  • Supplement: GlutaMax - Thermo Fisher Scientific Inc.
  • Supplement: Non-Essential Amino Acid Solution - Thermo Fisher Scientific Inc.
  • Supplement: 2-mercaptoethanol - Thermo Fisher Scientific Inc.
  • Supplement: Fibroblast growth factor 2, Peprotech - Thermo Fisher Scientific Inc.

Passage Method: Enzyme-free cell dissociation

External References

Source
Stem cell research Generation of a human ACTA1-tdTomato reporter iPSC line using CRISPR/Cas9 editing
Proceedings of the National Academy of Sciences of the United States of America Modeling human skeletal development using human pluripotent stem cells
Seminars in cell & developmental biology Developmental principles informing human pluripotent stem cell differentiation to cartilage and bone
Stem cell research Generation of a SOX9-tdTomato reporter human iPSC line, MCRIi001-A-2, using CRISPR/Cas9 editing
Computational and structural biotechnology journal Probing pluripotency gene regulatory networks with quantitative live cell imaging
Stem cell research CRISPR/Cas9 gene editing of a SOX9 reporter human iPSC line to produce two TRPV4 patient heterozygous missense mutant iPSC lines, MCRIi001-A-3 (TRPV4 p.F273L) and MCRIi001-A-4 (TRPV4 p.P799L)
Stem cell research Generation of a heterozygous COL2A1 (p.R989C) spondyloepiphyseal dysplasia congenita mutation iPSC line, MCRIi001-B, using CRISPR/Cas9 gene editing
hPSCreg MCRIi001-A-2
Cellosaurus CVCL_WU54
BioSamples SAMEA5873308
Wikidata Q95986993
Phenomics Australia Phenomics Australia