C6
MRIi001-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 | No disease was diagnosed.
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Karyotyping (Donor) |
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| Has the donor karyotype been analysed? |
No
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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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Donor Relations |
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| Other cell lines of this donor | |
External Databases (Donor) |
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| BioSamples | SAMEA104132876 |
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? | Yes |
| Do you (Depositor/Provider) hold the original Donor Consent Form? | Yes |
| Confirm that consent was obtained by a qualified professional | Yes |
| Has the donor agreed to be re-contacted? | Unknown |
| 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 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? | No information |
| Will the donor expect to receive financial benefit, beyond reasonable expenses, in return for donating the biosample? | No |
| 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? | Ethical Commission of the Faculty of Medicine (TUM) |
| Approval number | 2109/08 |
| 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? | Ethical Commission of the Faculty of Medicine (TUM) |
| Approval number | 2109/08 |
| 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? | Thermofisher |
hIPSC Derivation
General |
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| Source cell type |
Any skin fibroblast that is part of some dermis.
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Reprogramming method |
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| Vector type | Non-integrating |
| Vector | Sendai virus |
| 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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Other |
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| Derived under xeno-free conditions |
Unknown |
| Derived under GMP? |
No |
| Available as clinical grade? |
No |
Culture Conditions
| Surface coating | Matrigel/Geltrex |
| Passage method |
Enzyme-free cell dissociation
EDTA
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| CO2 Concentration | 5 % |
| Medium |
Essential 8™
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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? | Yes |
| 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 |
| TRA 1-81 |
Yes |
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| NANOG |
Yes |
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| OCT4 - POU class 5 homeobox 1 |
Yes |
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| SOX2 |
Yes |
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| c-MYC - MYC proto-oncogene, bHLH transcription factor |
Yes |
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| REX1 - RNA exonuclease 1 homolog |
Yes |
Morphology pictures
Representative colony of the C6 iPSC line. Scale bar, 100 µm.
Differentiation Potency
In vitro spontaneous differentiation
| Marker | Expressed |
| alpha fetoprotein |
Yes |
| SOX7 SRY-box transcription factor 7 |
Yes |
Protocol or reference
AFP_expression.png
Quantitative RT-PCR for the AFP gene expression representative of the endoderm germ layer upon spontaneous embryoid-body differentiation (D21 vs D0).
SOX7_expression.png
Quantitative RT-PCR for the SOX7 gene expression representative of the endoderm germ layer upon spontaneous embryoid-body differentiation (D21 vs D0).
In vitro spontaneous differentiation
| Marker | Expressed |
| ACTA2 – actin alpha 2, smooth muscle |
Yes |
| PECAM1 platelet and endothelial cell adhesion molecule 1 |
Yes |
Protocol or reference
ACTA2_expression.png
Quantitative RT-PCR for the ACTA2 gene expression representative of the mesoderm germ layer upon spontaneous embryoid-body differentiation (D21 vs D0).
CD31_expression.png
Quantitative RT-PCR for the CD31 (PECAM1) gene expression representative of the mesoderm germ layer upon spontaneous embryoid-body differentiation (D21 vs D0).
In vitro spontaneous differentiation
| Marker | Expressed |
| KRT14 keratin 14 |
Yes |
| TH tyrosine hydroxylase |
Yes |
Protocol or reference
KRT14_expression.png
Quantitative RT-PCR for the KRT14 gene expression representative of the ectoderm germ layer upon spontaneous embryoid-body differentiation (D21 vs D0).
TH_expression.png
Quantitative RT-PCR for the TH gene expression representative of the ectoderm germ layer upon spontaneous embryoid-body differentiation (D21 vs D0).
Microbiology / Virus Screening |
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| HIV 1 | Negative |
| Hepatitis B | Negative |
| Hepatitis C | Negative |
| Mycoplasma | Negative |
Genotyping
Karyotyping (Cell Line) |
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| Has the cell line karyotype been analysed? |
Yes
Normal
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Other Genotyping (Cell Line) |
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