D4-cB, M4C4, NP0053-5
The cell line is not validated yet.
UKKi006-A
General
Donor Information
General Donor Information |
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| Sex | female |
| Age of donor (at collection) | 60-64 |
| Ethnicity | Caucasian |
Phenotype and Disease related information (Donor) |
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| Diseases | No disease was diagnosed.
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| Disease associated phenotypes |
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| Family history | No |
| Is the medical history available upon request? | No |
| Is clinical information available? | No |
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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External Databases (Donor) |
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| BioSamples | SAMEA2825874 |
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. | |
| 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? | No |
| 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 |
| 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? | 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? | 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 |
| 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? | Yes |
| Has the donor consented to receive information discovered during use of donated embryo/tissue that has significant health implications for the donor? | 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 |
| Does the consent anticipate that the donor will be notified of results or outcomes of any research involving the donated samples or derived cells? | 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? | No |
| Does the consent permit the donor, upon withdrawal of consent, to stop delivery or use of information and data about 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? | RWTH Aachen |
| Approval number | |
| 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? | |
hIPSC Derivation
General |
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| Source cell type |
A mesenchymal stem cell that is part of the bone marrow.
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| Source cell origin |
The soft tissue that fills the cavities of bones.
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| Source cell type (free text) | Bone marrow-derived mesenchymal stromal cells (MSCs) |
| Age of donor (at collection) | 60-64 |
| Collected in | 2010 |
| Passage number reprogrammed | P4 |
Reprogramming method |
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| Vector type | Integrating |
| Vector | Virus (Retrovirus) |
| Genes | |
| Is the used vector excisable? |
No |
| Absence of reprogramming vector(s)? |
No |
| Reprogramming vectors silenced? |
Yes |
| Methods used |
RT-PCR
|
| Notes on reprogramming vector silencing | Retrovirally encoded reprogramming factors OSKM are not expressed in iPSCs as determined by RT-PCR |
| Files and images showing reprogramming vector expressed or silenced | |
| Vector map | |
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 | human ESC morphology |
| Derived under xeno-free conditions |
No |
| 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™
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| Has Rock inhibitor (Y27632) been used at passage previously with this cell line? | No |
| 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? | No |
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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| SSEA-4 |
Yes |
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| TRA 1-80 |
Yes |
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| TRA1-80 |
Yes |
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Morphology pictures
brightfield image of NP0053-5 iPSC colonies, day 2 post-thaw; 10x objective
brightfield image of NP0053-5 iPSC colony, day 2 post-thaw; 32x objective
Differentiation Potency
Microbiology / Virus Screening |
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| HIV 1 | Negative |
| HIV 2 | Negative |
| Hepatitis B | Negative |
| Hepatitis C | Negative |
| Mycoplasma | Negative |
Certificate of Analysis |
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| Is there a certificate of analysis available? |
Yes
Passage:
59
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Genotyping
Karyotyping (Cell Line) |
|
| Has the cell line karyotype been analysed? |
Yes
Deletion of the short arm and duplikation of the long arm of Chr20 were detected.
Karyotyping method:
Molecular karyotyping by SNP array
http:// |
Other Genotyping (Cell Line) |
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| Is there genome-wide genotyping or functional data available? |
Yes
Methylation profiling
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