SUSMi005-A
General
Donor Information
General Donor Information |
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| Sex | male |
| Ethnicity | White |
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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| Family history | positive |
| Is the medical history available upon request? | Yes, see Byers et al. 2011 |
| Is clinical information available? | Yes |
Karyotyping (Donor) |
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| Has the donor karyotype been analysed? |
Yes
46, XY
Karyotyping method:
G-Banding
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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 | SAMEA12279540 |
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. | Stanford University contracts office |
| Do you (Depositor/Provider) hold the original Donor Consent Form? | Yes |
| 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? | 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? | No |
| Does consent permit unforeseen future research, without further consent? | Yes |
| 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? | Yes |
| Does consent expressly permit storage of cells derived from the donated embryo/tissue for an unlimited time? | Yes |
| 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 |
| a public organisation? | Yes |
| a non-profit company? | Yes |
| a for-profit corporation? | 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? | Yes |
| Has the donor been informed that their donated biosample or derived cells may be tested for the presence of microbiological agents / pathogens? | Yes |
| 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 anticipate that the donor will be notified of results or outcomes of any research involving the donated samples or derived cells? | Yes |
| 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? | No |
| Does consent permit access to any other source of information about the clinical treatment or health of the donor? | 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? | Stanford University IRB |
| Approval number | IRB-15028 |
| 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? | Stanford University SCRO |
| Approval number | SCRO-212 |
| 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? | Addgene |
| 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 | |
Reprogramming method |
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| Vector type | Integrating |
| Vector | Virus (Retrovirus) |
| Genes | |
| Is the used vector excisable? |
Yes |
| Absence of reprogramming vector(s)? |
Yes |
| Reprogramming vectors silenced? |
Yes |
| Methods used |
RT-PCR
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| Notes on reprogramming vector silencing | See Byers et al Results: Generation of iPSCs |
Vector free reprogramming |
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Other |
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| Selection criteria for clones | morphology |
| Derived under xeno-free conditions |
No |
| Derived under GMP? |
No |
| Available as clinical grade? |
No |
Culture Conditions
| Surface coating | Gelatin | ||||||||||||||||||
| Feeder cells |
irradiated mouse embryonic fibroblast |
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| Passage method | Mechanically | ||||||||||||||||||
| O2 Concentration | 20 % | ||||||||||||||||||
| CO2 Concentration | 5 % | ||||||||||||||||||
| Medium |
Other medium:
Base medium: DMEM/F12
Main protein source: Knock-out serum replacement Serum concentration: 20 % Supplements
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| Has Rock inhibitor (Y27632) been used at passage previously with this cell line? | Yes |
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| Has Rock inhibitor (Y27632) been used at cryo previously with this cell line? | No |
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| 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 1-60 |
Yes |
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| TRA 1-81 |
Yes |
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| Alkaline Phosphatase |
Yes |
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| NANOG |
Yes |
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| SSEA-1 |
Yes |
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| SSEA-3 |
Yes |
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| SSEA-4 |
Yes |
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Score:
| Marker | Present | Absent |
| mCpG | ||
| OCT4 |
Morphology pictures
Byers 2011.pdf
Byers 2011
The NANOG and OCT4 promoters were hypermethylated in the untransduced donor fibroblasts and hypomethylated in the reprogrammed iPSCs and H9 hESCs (Fig. S2A). Imprinted genes
maintained their differentially-methylated state in reprogrammed
colonies, confirming iPSC epigenetic stability and suggesting reduced tumorigenic risk and complete reprogramming (Fig. S2B). Likewise, iPSC telomerase activity was comparable to hESCs and was significantly elevated compared to untransduced fibroblasts.
Differentiation Potency
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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