LEIi004-A
General
Cell Line |
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| hPSCreg name | LEIi004-A |
| Cite as: | LEIi004-A (RRID:CVCL_VE61) |
| Cell line type | Human induced pluripotent stem cell (hiPSC) |
| Similar lines |
LUMCi056-A-1 (LUMC0128iCRB01 heterozygote CRISPR corrected isogenic clone 02, iso02LUMC0128iCRB01) Donor's gene variants: CRB1, CRB1 Donor diseases: Retinitis Pigmentosa UNEWi002-A (UNEW002Ai, PRPF31 AW) Donor's gene variants: PRPF31, PRPF31 Donor diseases: Retinitis pigmentosa ESi077-A (CABi001-A, PRPF31-MiPS4F3) Donor's gene variants: PRPF31, PRPF31 Donor diseases: Retinitis Pigmentosa LUMCi055-A (CRB1 patient 117 compound heterozygous 2983G>T p.(Glu995*) c.1892A>G, p.(Tyr631Cys), LUMC0117iCRB01) Donor's gene variants: CRB1, CRB1 Donor diseases: Retinitis Pigmentosa LUMCi056-A (CRB1 patient 128 compound heterozygous c.2843G>A p.(Cys948Tyr) and c.3122T>C p.(Met1041Thr), LUMC0128iCRB01) Donor's gene variants: CRB1, CRB1 Donor diseases: Retinitis Pigmentosa |
| Last update | 22nd May 2019 |
| User feedback | |
Provider |
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| Generator | Lions Eye Institute (LEI) |
| Owner | Lions Eye Institute (LEI) |
| Derivation country | Australia |
External Databases |
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| BioSamples | SAMEA104626352 |
| Cellosaurus | CVCL_VE61 |
| Wikidata | Q54902437 |
General Information |
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| Publications |
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| * Is the cell line readily obtainable for third parties? |
No |
| Subclones | |
Donor Information
General Donor Information |
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| Sex | female |
| Ethnicity | Caucasian |
Phenotype and Disease related information (Donor) |
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| Diseases | A disease was diagnosed.
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| Disease associated phenotypes |
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| Family history | RCD , CLN3 mutation |
| Is the medical history available upon request? | YES |
| Is clinical information available? | YES |
Karyotyping (Donor) |
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| Has the donor karyotype been analysed? |
Yes
Karyotyping method:
Molecular karyotyping by SNP array
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Other Genotyping (Donor) |
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| Is there genome-wide genotyping or functional data available? |
No
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External Databases (Donor) |
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| BioSamples | SAMEA104626353 |
Ethics
| Has informed consent been obtained from the donor of the embryo/tissue from which the pluripotent stem cells have been derived? | Yes |
| Was the consent voluntarily given? | Yes |
| Has the donor been informed that participation will not directly influence their personal treatment? | Yes |
| Can you provide us with a copy of the Donor Information Sheet provided to the donor? | No |
| Provide contact information of the holder of the original Donor Information Sheet: | fredchen@lei.org.au |
| Do you (Depositor/Provider) hold the original Donor Consent Form? | No |
| If you do not hold the Donor Consent Form, do you know who does? | Yes |
| Contact information / weblink | fredchen@lei.org.au |
| Alternatives to consent | |
| Alternative consent approval number | |
| Confirm that consent was obtained by a qualified professional | Yes |
| Has the donor agreed to be re-contacted? | Yes |
| Has the donor been informed about how her/his data will be protected? | Yes |
| Please indicate whether the data associated with the donated material has been pseudonymised or anonymised. | pseudonymised |
| Does consent explicitly allow the derivation of pluripotent stem cells? | No |
| Does consent expressly prevent the derivation of pluripotent stem cells? | Yes |
| Does consent pertain to a specific research project? | Yes |
| Details on restriction to research project | The Western Australia Retinal Degeneration (WARD) study |
| Does consent permit unforeseen future research, without further consent? | Yes |
| Does the consent permit uses of donated embryo/tissue or derived cell line intended for clinical treatment or human applications? | Yes |
| Does consent expressly prevent development of commercial products? | No |
| Does consent expressly prevent financial gain from any use of the donated embryo/tissue, including any product made from it? | No |
| Does consent expressly permit storage of donated embryo/tissue for an unlimited time? | No |
| Does consent expressly permit storage of cells derived from the donated embryo/tissue for an unlimited time? | No |
| Does consent prevent the DONATED BIOSAMPLE from being made available to researchers anywhere in the world? | No |
| Does consent prevent CELLS DERIVED FROM THE DONATED BIOSAMPLE from being made available to researchers anywhere in the world? | No |
Does consent permit research by | |
| an academic institution? | Yes |
| Does consent expressly permit collection of genetic information? | Yes |
| Does consent expressly permit storage of genetic information? | Yes |
| Does consent prevent dissemination of genetic information? | Yes |
| Has the donor been informed that their donated biosample or derived cells may be tested for the presence of microbiological agents / pathogens? | Yes |
| How may genetic information associated with the cell line be accessed? | Controlled Access |
| Will the donor expect to receive financial benefit, beyond reasonable expenses, in return for donating the biosample? | No |
| Does the consent anticipate that the donor will be notified of results or outcomes of any research involving the donated samples or derived cells? | Yes |
| Does the consent permit the donor, upon withdrawal of consent, to stop the use of the derived cell line(s) that have already been created from donated samples? | Yes |
| Does the consent permit the donor, upon withdrawal of consent, to stop delivery or use of information and data about the donor? | Yes |
| Does consent permit access to medical records of the donor? | Yes |
| Please describe how access is provided: | Lions Eye Institute |
| Does consent permit access to any other source of information about the clinical treatment or health of the donor? | Yes |
| Contact data, institution, or website: | Lions Eye Institute |
| Has a favourable opinion been obtained from a research ethics committee, or other ethics review panel, in relation to the Research Protocol including the consent provisions? | Yes |
| Name of accrediting authority involved? | UWA Human Research Ethics Committee |
| Approval number | RA/4/1/7916 |
| Has a favourable opinion been obtained from a research ethics committee, or other ethics review panel, in relation to the PROPOSED PROJECT, involving use of donated embryo/tissue or derived cells? | Yes |
| Name of accrediting authority involved? | UWA Human Research Ethics Committee |
| Approval number | RA/4/1/7916 |
| Do you have obligations to third parties in regard to the use of the cell line? | No |
| Please describe: | |
| Are you aware of any further constraints on the use of the donated embryo/tissue or derived cells? | No |
| Further constraints on use | |
| Is there an MTA available for the cell line? | No |
| For generation of the cell line, who was the supplier of any recombined DNA vectors or commercial kits used? | SBI Cat#SC900A-1 |
| Are you aware of any constraints on the use or distribution of the cell line from the owner or any parties identified in the query above? | No |
| Constraints for use or distribution | |
hIPSC Derivation
General |
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| Source cell type |
A connective tissue cell which secretes an extracellular matrix rich in collagen and other macromolecules. Flattened and irregular in outline with branching processes; appear fusiform or spindle-shaped.; These cells may be vimentin-positive, fibronectin-positive, fsp1-positive, MMP-1-positive, collagen I-positive, collagen III-positive, and alpha-SMA-negative.
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| Passage number reprogrammed | 7 |
Reprogramming method |
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| Vector type | Non-integrating |
| Vector | Episomal |
| Is reprogramming vector detectable? |
Yes |
| Methods used |
PCR
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| Files and images showing reprogramming vector expressed or silenced | |
Vector free reprogramming |
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| Type of used vector free reprogramming factor(s) |
None
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Other |
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| Selection criteria for clones | Morphology |
| Derived under xeno-free conditions |
Yes |
| Derived under GMP? |
Yes |
| Available as clinical grade? |
Yes |
Culture Conditions
| Surface coating | Matrigel/Geltrex |
| Feeder cells |
No |
| Passage method |
Enzyme-free cell dissociation
EDTA
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| O2 Concentration | 20 % |
| CO2 Concentration | 5 % |
| Medium |
mTeSR™ 1
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| Has Rock inhibitor (Y27632) been used at passage previously with this cell line? | Yes |
| Has Rock inhibitor (Y27632) been used at cryo previously with this cell line? | No |
| Has Rock inhibitor (Y27632) been used at thaw previously with this cell line? | Yes |
Characterisation
Analysis of Undifferentiated Cells
| Marker | Expressed | Immunostaining | RT-PCR | Flow Cytometry | Enzymatic Assay | Expression Profiles |
| NANOG |
Yes |
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| POU5F1 (OCT-4) |
Yes |
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| SOX2 |
Yes |
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| SSEA-4 |
Yes |
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Score:
| Marker | Present | Absent |
| mCpG | ||
| OCT4 |
Self-renewal
Positive
Endoderm
Positive
Mesoderm
Positive
Ectoderm score
Positive
Differentiation Potency
Genotyping
Karyotyping (Cell Line) |
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| Has the cell line karyotype been analysed? |
Yes
46+XX
Passage number: 25
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Other Genotyping (Cell Line) |
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