INSAi003-A
General
Cell Line |
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| hPSCreg name | INSAi003-A |
| Cite as: | INSAi003-A |
| Cell line type | Human induced pluripotent stem cell (hiPSC) |
| Similar lines | No similar lines found. |
| Last update | 7th June 2024 |
| User feedback | |
Provider |
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| Generator | Instituto Nacional de Saude Ricardo Jorge (INSA) |
| Owner | Instituto Nacional de Saude Ricardo Jorge (INSA) |
| Derivation country | Portugal |
External Databases |
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| BioSamples | SAMEA115474367 |
General Information |
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| * Is the cell line readily obtainable for third parties? |
No |
Donor Information
General Donor Information |
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| Sex | female |
Phenotype and Disease related information (Donor) |
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| Diseases | A disease was diagnosed.
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| Disease associated phenotypes |
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Karyotyping (Donor) |
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| Has the donor karyotype been analysed? |
Unknown
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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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External Databases (Donor) |
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| BioSamples | SAMEA115474368 |
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? | No |
| If you do not hold the Donor Consent Form, do you know who does? | Yes |
| Please provide the contact information | geneticbiobank@gaslini.org |
| Alternatives to consent are available? | No |
| Is there other documentation provided to the donor for consenting purposes? | No |
| Confirm that consent was obtained by a qualified professional | Yes |
| Has the donor agreed to be re-contacted? | Unknown |
| 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? | No |
| Does consent expressly prevent the derivation of pluripotent stem cells? | No |
| Does consent pertain to a specific research project? | No |
| Does the consent permit uses of donated embryo/tissue or derived cell line intended for clinical treatment or human applications? | 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 expressly permit storage of donated embryo/tissue for an unlimited time? | No |
| Does consent expressly permit storage of cells derived from the donated embryo/tissue for an unlimited time? | 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 |
| 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 anticipate that the donor will be notified of results or outcomes of any research involving the donated samples or derived cells? | Yes |
| 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? | Comissão de Ética para a Saúde do INSA (CES-INSA); Decreto-Lei n.º 80/2018 |
| Approval number | |
| 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? | Comissão de Ética para a Saúde do INSA (CES-INSA); Decreto-Lei n.º 80/2018 |
| Approval number | |
| Do you have obligations to third parties in regard to the use of the cell line? | No |
| Is there an MTA available for the cell line? | Yes |
| For generation of the cell line, who was the supplier of any recombined DNA vectors or commercial kits used? | Epi5™ Episomal iPSC Reprogramming Kit (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 line name | Skin fibroblasts |
| Source cell type | |
| Source cell origin |
An organ that constitutes the external surface of the body. It consists of the epidermis, dermis, and skin appendages.
Synonyms
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| Passage number reprogrammed | 6 |
Reprogramming method |
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| Vector type | Non-integrating |
| Vector | Episomal |
| Genes | |
| Is reprogramming vector detectable? |
Yes |
| Methods used |
PCR
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| Notes on reprogramming vector detection | Endpoint PCR detection of transgenes oriP and EBNA-1 using the primers described in the |
| Files and images showing reprogramming vector expressed or silenced | |
Vector free reprogramming |
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| Type of used vector free reprogramming factor(s) |
None
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Other |
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| Selection criteria for clones | The clones were initially isolated by manual picking, according to their tipical morphology, then EDTA passage. The clones were selected based on the expression of pluripotency markers and normal kariotype. |
| Derived under xeno-free conditions |
Yes |
| Derived under GMP? |
No |
| Available as clinical grade? |
No |
Culture Conditions
| Surface coating | Vitronectin |
| Feeder cells |
No |
| Passage method |
Enzyme-free cell dissociation
EDTA
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| O2 Concentration | 20 % |
| CO2 Concentration | 5 % |
| Medium | Essential 8™ Flex |
| 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 |
| NANOG |
Yes |
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| POU5F1 (OCT-4) |
Yes |
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| SOX2 |
Yes |
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| TRA 1-60 |
Yes |
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Score:
| Marker | Present | Absent |
| mCpG | ||
| OCT4 |
Differentiation Potency
In vitro directed differentiation
| Marker | Expressed |
| SOX17 |
Yes |
Protocol or reference
Protocol - iPSCs diff three germ layers.pdf
Protocol of differentiation into the three germ layers
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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