iOP118
KCLi001-A
General
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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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
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External Databases (Donor) |
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| BioSamples | SAMEA4932835 |
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. | ethikkommission@i-med.ac.at |
| 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 | ethikkommission@i-med.ac.at |
| 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? | Yes |
| Does consent expressly prevent the derivation of pluripotent stem cells? | No |
| Details on restriction to research project | |
| 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 |
| 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? | Ethics Committee of the Medical University of Innsbruck, Austria |
| Approval number | AN2016-0260 |
| 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? | Ethics Committee of the Medical University of Innsbruck, Austria |
| Approval number | AN2016-0260 |
| 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 |
An epidermal cell which synthesizes keratin and undergoes a characteristic change as it moves upward from the basal layers of the epidermis to the cornified (horny) layer of the skin. Successive stages of differentiation of the keratinocytes forming the epidermal layers are basal cell, spinous or prickle cell, and the granular cell.; Keratinocytes are reportedly CDw210a-negative, CDw210b-positive, CD281-positive, CD282-positive, CD285-positive, IL22Ra1-positive, Human keratinocytes are reportedly capable of secreting BD-2, BD-3, hCAP-18, CXCL1, CXCL5, CXCL8, elafin, MMP-3, NGAL, PDGF-A, S100A7, S100A8, and S100A9. Transcription factors: STAT3-positive.
Synonyms
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| Source cell origin |
Any portion of the organ that covers that body and consists of a layer of epidermis and a layer of dermis.
Synonyms
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| Passage number reprogrammed | 2 |
Reprogramming method |
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| Vector type | Non-integrating |
| Vector | Sendai virus |
| Genes | |
| Is reprogramming vector detectable? |
Yes |
| Methods used |
RT-PCR
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| Notes on reprogramming vector detection | After ten passages, the elimination of the SeV vectors was confirmed in the KCLi001-A cell line by RT-PCR using specific primers |
| 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 | Three weeks post-transduction colonies with ES-like morphology appeared and were selected to establish feeder-free iPSC clones |
| Derived under xeno-free conditions |
Yes |
| Derived under GMP? |
No |
| Available as clinical grade? |
No |
Culture Conditions
| Surface coating | Matrigel/Geltrex |
| Feeder cells |
Irradiated Human Foreskin Fibroblasts (HFF) |
| Passage method |
Enzyme-free cell dissociation
Gentle Cell Dissociation Reagent
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| O2 Concentration | 5 % |
| CO2 Concentration | 5 % |
| Medium |
mTeSR™ 2
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| Has Rock inhibitor (Y27632) been used at passage previously with this cell line? | No |
| 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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| TRA 1-60 |
Yes |
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| TRA 1-81 |
Yes |
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| Alkaline Phosphatase |
Yes |
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Differentiation Potency
In vivo teratoma
In vitro spontaneous differentiation
In vitro directed differentiation
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
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Other Genotyping (Cell Line) |
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