iPSC_BrS9_FB_C7
BBANTWi006-A
General
Donor Information
General Donor Information |
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| Sex | male |
| Ethnicity | White Belgian |
Phenotype and Disease related information (Donor) |
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| Diseases | A disease was diagnosed.
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| Disease associated phenotypes |
|
| Family history | yes |
| Is the medical history available upon request? | available upon request |
| Is clinical information available? | available upon request |
Karyotyping (Donor) |
|
| Has the donor karyotype been analysed? |
Yes
46, XY. No clinically significant abnormalities observed.
Karyotyping method:
Molecular karyotyping by SNP array
http:// |
Other Genotyping (Donor) |
|
| Is there genome-wide genotyping or functional data available? |
Yes
SNP typing array
Skin fibroblast DNA sample was genotyped with Illumina's HumanCytoSNP-12 v2.1. No clinically significant CNVs observed. |
External Databases (Donor) |
|
| BioSamples | SAMEA9372405 |
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? | 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? | Yes |
| Does consent expressly prevent the derivation of pluripotent stem cells? | No |
| Does consent pertain to a specific research project? | Yes |
| Details on restriction to research project | Study of inherited cardiovascular diseases |
| Does consent permit unforeseen future research, without further consent? | No |
| 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 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? | No |
| Has the donor been informed that their donated biosample or derived cells may be tested for the presence of microbiological agents / pathogens? | No |
| Has the donor consented to receive information discovered during use of donated embryo/tissue that has significant health implications for the donor? | 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 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: | Through the treating physician |
| 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? | UZA ethics committee |
| Approval number | EC 16/49/537 |
| 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? | UZA ethics committee |
| Approval number | EC 16/49/537 |
| 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 |
| 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? | CytoTune™-iPS 2.0 Sendai Reprogramming Kit from Invitrogen / ThermoFisher 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 | |
| Source cell origin | |
| Source cell type (free text) | Skin biopsy was taken on inner side of upper arm |
| Passage number reprogrammed | 5 |
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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| Notes on reprogramming vector detection | SeV not detected on passage 11 |
Vector free reprogramming |
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Other |
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| Selection criteria for clones | Clones were picked based on morphology (round, flat colonies with smooth edges and tightly packed cells) for 5 rounds and subsequently passaged using EDTA for 5 more rounds. Only clones with nice morphology and no differentiation after these 10 passaging rounds were selected. |
| Derived under xeno-free conditions |
No |
| Derived under GMP? |
Unknown |
| Available as clinical grade? |
No |
Culture Conditions
| Surface coating | Matrigel/Geltrex |
| Feeder cells |
No |
| Passage method |
Enzyme-free cell dissociation
EDTA
|
| O2 Concentration | 5 % |
| CO2 Concentration | 5 % |
| Medium |
Essential 8™ Flex
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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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| TRA 1-60 |
Yes |
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| TRA 1-81 |
Yes |
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| NANOG |
Yes |
Differentiation Potency
In vitro spontaneous differentiation
| Marker | Expressed |
| SOX17 |
Yes |
| CXCR4 |
Yes |
In vitro spontaneous differentiation
In vitro directed differentiation
| Marker | Expressed |
| SCN5A |
Yes |
| Troponin I, cardiac muscle |
Yes |
| NKX2-5 |
Yes |
| alpha-actinin 2 |
Yes |
| MYL2 |
Yes |
Microbiology / Virus Screening |
|
| Mycoplasma | Negative |
Genotyping
Karyotyping (Cell Line) |
|
| Has the cell line karyotype been analysed? |
Yes
46, XY. No clinically significant abnormalities observed.
Passage number: 11
Karyotyping method:
Molecular karyotyping by SNP array
http:// |
Other Genotyping (Cell Line) |
|
| Is there genome-wide genotyping or functional data available? |
Yes
SNP typing array
Genotyped with Illumina's HumanCytoSNP-12 v2.1. No clinically significant CNVs observed. |
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