MCRIi001-A-4

Not yet reviewed by Australian Stem Cell Registry

TRPV4 p.P799L, SOX9tdTom TRPV4 p.P799L, PB001-SOX9tdTOM-TRPV4pP799L, MCRIi001-A-2-TRPV4 p.P799L

Details

Tissue & Disease as reported

Disease as reported (Genomic Modifications)
metatropic dysplasia (Isogenic modification)

Genetic Information

Genotype Locus
TRPV4
Polymorphism
Heterozygous: unspecified_reference_TRPV4:c.2396C>T(p.Phe799Leu)
Modifications
Isogenic modification

Associated Publications

Journal Year Article
Methods in molecular biology (Clifton, N.J.) 2023 Chondrogenic Differentiation of Human-Induced 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 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 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).

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

Heterozygous: unspecified_reference_TRPV4:c.2396C>T(p.Phe799Leu)

Heterozygous TRPV4 (c.2396C>T; p.Phe799Leu) variant was introduced to TRPV4 to generate a model of Metatropic Dysplasia. Synonymous changes were also introduced.

Cytoband: 12q24.11
Mutation Type: Isogenic modification
Delivery Method: CRISPR/Cas9

Quality Assurance

Genomic Characterisation

Passage Number: 5

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

Karyotype Method: Molecular karyotyping by SNP array

Summary: Molecular karyotyping by SNP array (Illumina Infinium CoreExome-24 v1.1; 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
NANOG Immunostaining
POU5F1 (OCT-4) Immunostaining
CD9 Flow cytometry
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 vivo teratoma

Assessed by: Immunostaining

Markers: HNF4A

Mesoderm

In vivo teratoma

Assessed by: Immunostaining

Markers: COL2A1

Ectoderm

In vivo teratoma

Assessed by: Immunostaining

Markers: MAP2

Growth Characteristics

Culture Medium and Growth Conditions

CO2 concentration: 5%

O2 concentration: Unavailable

Medium Items:

  • Supplement: Fibroblast growth factor 2, Peprotech - Thermo Fisher Scientific Inc.
  • Base Coat: Mouse embryonic fibroblasts
  • Base Medium: DMEM/F-12 (Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12) - Thermo Fisher Scientific Inc.
  • Serum: Knock-out Serum Replacement - Thermo Fisher Scientific Inc.
  • Supplement: Non-Essential Amino Acids - Thermo Fisher Scientific Inc.
  • Supplement: GlutaMAX - Thermo Fisher Scientific Inc.
  • Supplement: 2-mercaptoethanol - Thermo Fisher Scientific Inc.

Passage Method: Enzyme-free cell dissociation

External References

Source
Methods in molecular biology (Clifton, N.J.) Chondrogenic Differentiation of Human-Induced Pluripotent Stem Cells
Seminars in cell & developmental biology Developmental principles informing human pluripotent stem cell differentiation to cartilage and bone
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 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).
hPSCreg MCRIi001-A-4
Cellosaurus CVCL_A9Z4
Wikidata Q102114509
Phenomics Australia Phenomics Australia