RYR1-15377-R1B
HPIi009-A
General
Cell Line |
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| hPSCreg name | HPIi009-A |
| Cite as: | HPIi009-A (RRID:CVCL_D6KI) |
| Alternative name(s) |
RYR1-15377-R1B
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| Cell line type | Human induced pluripotent stem cell (hiPSC) |
| Similar lines |
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| Last update | 22nd March 2024 |
| Notes | iPSC line derived from EBV-immortalised LCLs (Genethon ID 5-15377). Line has a heterozygous variant in the RYR1 gene: NM_000540.3: c.4038C>A (p.Asn1346Lys). |
| User feedback | |
Provider |
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| Generator | Harry Perkins Institute of Medical Research (HPI) |
| Owner | Harry Perkins Institute of Medical Research (HPI) |
| Distributors | |
| Derivation country | Australia |
External Databases |
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| BioSamples | SAMEA115133464 |
| Cellosaurus | CVCL_D6KI |
| Wikidata | Q127381784 |
General Information |
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| Publications | |
| * Is the cell line readily obtainable for third parties? |
Yes Research use: allowed
Clinical use: not allowed
Commercial use: not allowed
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Donor Information
General Donor Information |
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| Sex | male |
| Age of donor (at collection) | 25-29 |
| Ethnicity | Caucasian |
Phenotype and Disease related information (Donor) |
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| Diseases | A disease was diagnosed.
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Karyotyping (Donor) |
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| Has the donor karyotype been analysed? |
Yes
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Other Genotyping (Donor) |
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| Is there genome-wide genotyping or functional data available? |
Yes
Phased long-read sequencing of RYR1 gene
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External Databases (Donor) |
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| BioSamples | SAMEA115133471 |
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 |
| Please provide contact information of the holder of the original Donor Information Sheet. | Norma B. Romero, nb.romero@institut-myologie.org |
| 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 |
| Please provide the contact information | Norma B. Romero, nb.romero@institut-myologie.org |
| Confirm that consent was obtained by a qualified professional | Yes |
| Has the donor agreed to be re-contacted? | Unknown |
| 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? | No |
| Does consent pertain to a specific research project? | No |
| Does consent permit unforeseen future research, without further consent? | No |
| Does consent expressly permit storage of cells derived from the donated embryo/tissue for an unlimited time? | Yes |
| Does consent prevent CELLS DERIVED FROM THE DONATED BIOSAMPLE from being made available to researchers anywhere in the world? | No |
| 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 |
| 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? | Comité de Protection des Personnes |
| Approval number | Est IV DC-2012-1693 |
| 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? | University of Western Australia Human Research Ethics Committee |
| Approval number | RA/4/20/1008 |
| Do you have obligations to third parties in regard to the use of the cell line? | No |
| Are you aware of any further constraints on the use of the donated embryo/tissue or derived cells? | No |
| For generation of the cell line, who was the supplier of any recombined DNA vectors or commercial kits used? | Thermo Fisher Scientific |
| 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 |
hIPSC Derivation
General |
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| Source cell type |
Human cell line from tissue infected with Epstein-Barr virus, resembling a lymphoblast.
Synonyms
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| Source cell origin |
A liquid tissue; its major function is to transport oxygen throughout the body. It also supplies the tissues with nutrients, removes waste products, and contains various components of the immune system defending the body against infection. Several hormones also travel in the blood.
Synonyms
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| Source cell type (free text) | iPSCs were reprogrammed from EBV-immortalised LCLs banked at Genethon. EBV clearance was validated for these lines. |
| Age of donor (at collection) | 25-29 |
| Passage number reprogrammed | 5 |
Reprogramming method |
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| Vector type | Non-integrating |
| Vector | Sendai virus |
| Genes | |
| Is reprogramming vector detectable? |
No |
| Methods used |
RT-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 | Emerging clones (good morphology) were manually picked from 6-well plates into individual 24-wells. The 'best' clones (based on morphology and lack of spontaneous differentiation) were then manually cleaned and picked again into 12-wells. Individual clones were then manually cleaned (to remove any spontaneous differentiation) before Versene passaging into a 6-well. From this point on, clones were passaged using Versene at a 1:6 to 1:20 split ratio. |
| Derived under xeno-free conditions |
No |
| Derived under GMP? |
No |
| Available as clinical grade? |
No |
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™ Plus
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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 |
| POU5F1 (OCT-4) |
Yes |
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| SOX2 |
Yes |
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| SSEA-4 |
Yes |
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| TRA 1-60 |
Yes |
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| NANOG |
Yes |
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| CRIPTO |
Yes |
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Morphology pictures
Differentiation Potency
In vitro directed differentiation
| Marker | Expressed |
| GATA4 |
Yes |
| SOX17 |
Yes |
Morphology
Immunofluorescence for SOX17 in HPIi009-A, another line HPIi008-A and control SCTi003-A, following directed endoderm differentiation (STEMdiff).
In vitro directed differentiation
| Marker | Expressed |
| TBXT |
Yes |
| TBX6 |
Yes |
Morphology
Immunofluorescence for Brachyury (TBXT) in HPIi009-A, other line HPIi008-A and control SCTi003-A, following directed mesoderm differentiation (STEMdiff).
In vitro directed differentiation
| Marker | Expressed |
| OTX2 |
Yes |
| PAX6 |
Yes |
Morphology
Immunofluorescence for OTX2 in HPIi009-A, other line HPIi008-A and control SCTi003-A, following directed ectoderm differentiation (STEMdiff).
Microbiology / Virus Screening |
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| Mycoplasma | Negative |
Genotyping
Karyotyping (Cell Line) |
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| Has the cell line karyotype been analysed? |
Yes
46,XY
Passage number: 13
Karyotyping method:
G-Banding
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Other Genotyping (Cell Line) |
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