IDISi001-A
General
Donor Information
General Donor Information |
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| Sex | male |
| 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? |
Unknown
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External Databases (Donor) |
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| BioSamples | SAMEA4675696 |
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. | Dr. Francisco Campos Pérez (francisco.campos.perez@sergas.es) |
| 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 | Dr. Francisco Campos Pérez (francisco.campos.perez@sergas.es) |
| Alternatives to consent | |
| Alternative consent approval number | |
| 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 |
| Details on restriction to research project | |
| Does consent expressly permit storage of donated embryo/tissue for an unlimited time? | Yes |
| 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? | Yes |
| How may genetic information associated with the cell line be accessed? | No information |
| Will the donor expect to receive financial benefit, beyond reasonable expenses, in return for donating the biosample? | No |
| Please describe how access is provided: | |
| Contact data, institution, or website: | |
| 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? | Commission on Guarantees for the Donation and Use of Human Cells and Tissues of Instituto de Salud Carlos III |
| Approval number | 428-346-1 |
| 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? | Commission on Guarantees for the Donation and Use of Human Cells and Tissues of Instituto de Salud Carlos III |
| Approval number | 428-346-1 |
| Do you have obligations to third parties in regard to the use of the cell line? | No |
| Please describe: | |
| Further constraints on use | |
| For generation of the cell line, who was the supplier of any recombined DNA vectors or commercial kits used? | |
| Constraints for use or distribution |
hIPSC Derivation
General |
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| Source cell type |
A leukocyte with a single non-segmented nucleus in the mature form.
Synonyms
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Reprogramming method |
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| Vector type | Non-integrating |
| Vector | Sendai virus |
| Genes | |
| Is reprogramming vector detectable? |
Yes |
| Methods used |
PCR
|
| Notes on reprogramming vector detection | Detection of the DNA vector by PCR of a sample of PBMCs 24h after infection. Absence of the vector was checked by PCR of several clones of the new line at passage 5 |
| Files and images showing reprogramming vector expressed or silenced | |
Vector free reprogramming |
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Other |
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| Selection criteria for clones | For the first and second passages, colonies were selected and pickedwith the help of an Origio Stripper (Origio)with Stripper tips (Origio) of 175 μm in diameter. Later passages were performed mechanically, using cell lifters. |
| Derived under xeno-free conditions |
Unknown |
| Derived under GMP? |
Unknown |
| Available as clinical grade? |
Unknown |
Culture Conditions
| Surface coating | Matrigel/Geltrex | ||||||
| Feeder cells |
No |
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| Passage method | Mechanically | ||||||
| CO2 Concentration | 5 % | ||||||
| Medium |
mTeSR™ 1
Supplements
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| Has Rock inhibitor (Y27632) been used at passage previously with this cell line? | No |
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| Has Rock inhibitor (Y27632) been used at cryo previously with this cell line? | No |
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| Has Rock inhibitor (Y27632) been used at thaw previously with this cell line? | No |
Characterisation
Analysis of Undifferentiated Cells
| Marker | Expressed | Immunostaining | RT-PCR | Flow Cytometry | Enzymatic Assay | Expression Profiles |
| Alkaline Phosphatase |
Yes |
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| 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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| TRA 1-60 |
Yes |
Score:
| Marker | Present | Absent |
| mCpG | ||
| OCT4 |
Differentiation Potency
In vitro spontaneous differentiation
| Marker | Expressed |
| FOXA2 |
Yes |
Morphology
EBs 05-05-17 HF_Image018.tif
FOXA2 (Green) + DAPI (Blue)
In vitro spontaneous differentiation
| Marker | Expressed |
| Vimentin |
Unknown |
Morphology
EBs 05-05-17 HF_Image032.tif
Vimentin (Green) + DAPI (Blue)
EBs 05-05-17 HF_Image028.tif
Vimentin (Green) + DAPI (Blue)
In vitro spontaneous differentiation
| Marker | Expressed |
| NES |
Unknown |
Morphology
EBs 19-05-17_Image012.tif
Nestin (Green) + DAPI (Blue)
EBs 19-05-17_Image025.tif
Nestin (Green) + DAPI (Blue)
Microbiology / Virus Screening |
|
| Mycoplasma | Negative |
Genotyping
Karyotyping (Cell Line) |
|
| Has the cell line karyotype been analysed? |
Yes
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Other Genotyping (Cell Line) |
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