iPSC_FB_M45-50_C15
BBANTWi001-B
General#
Cell Line |
|
| hPSCreg Name | BBANTWi001-B |
| Alternative name(s) |
iPSC_FB_M45-50_C15
|
| Cell line type | Human induced pluripotent stem cell (hiPSC) |
| Last update | 25th June 2021 |
| User feedback | |
Provider |
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| Generator | Biobank Antwerpen (BBANTW) |
| Owner | Center of Medical Genetics Antwerp (CMGANT) |
| Derivation country | Belgium |
External Databases |
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| Cellosaurus | CVCL_ZX26 |
| BioSamples | SAMEA9269224 |
| CLO | CLO_0103348 |
General Information |
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| * Is the cell line readily obtainable for third parties? |
Yes Research: allowed
Clinical: not allowed
Commercial: not allowed
|
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 | No disease was diagnosed.
|
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. |
Donor Relations |
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| Other cell lines of this donor | |
External Databases (Donor) |
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| BioSamples | SAMEA9269223 |
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 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 18/12/166 |
| 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 18/12/166 |
| 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 |
hIPSC Derivation#
General |
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| Source cell type |
skin fibroblast |
| Source cell origin |
upper arm |
| Source cell type (free text) | Skin biopsy was taken on inner side of upper arm |
| Passage number reprogrammed | 6 |
Reprogramming method |
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| Vector type | Non-integrating |
| Vector | Sendai virus |
| Is reprogramming vector detectable? |
No |
| Methods used |
RT-PCR
|
| 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
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| O2 Concentration | 21 % |
| 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 | FACS | 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 |
| CXCR4 |
Yes |
| GATA6 |
Yes |
In vitro directed differentiation
| Marker | Expressed |
| Troponin I, cardiac muscle |
Yes |
| alpha-actinin 2 |
Yes |
| SCN5A |
Yes |
| NKX2-5 |
Yes |
| MYL2 |
Yes |
In vitro spontaneous differentiation
| Marker | Expressed |
| PAX6 |
Yes |
| HES5 |
Yes |
| NEUROD1 |
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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