PD30-5-16
ICGi034-B
General
Donor Information
General Donor Information |
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| Sex | female |
| Age of donor (at collection) | 55-59 |
| Ethnicity | Caucasian |
Phenotype and Disease related information (Donor) |
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| Diseases | A disease was diagnosed.
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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? |
Yes
Exome sequencing
NCBI SRR accession: SAMN22788974, BioProject PRJNA563295 Detailed analysis of the clinical exome sequencing data of the patient's PBMCs has revealed a mutation in the GBA gene (N370S). |
Donor Relations |
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| Other cell lines of this donor | |
| All cell lines of this donor's relatives | |
External Databases (Donor) |
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| BioSamples | SAMEA10450937 |
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 |
| 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? | Yes |
| 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? | Yes |
| Does consent expressly prevent financial gain from any use of the donated embryo/tissue, including any product made from it? | Yes |
| 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? | No |
| a non-profit company? | No |
| a for-profit corporation? | No |
| 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 |
| 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? | 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? | FSBI Federal Neurosurgical Center |
| Approval number | protocol number 1, 14/03/2017 |
| 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 |
A peripheral blood cell with a single nucleus. This category includes lymphocytes and monocytes.
Synonyms
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| Source cell origin |
Blood drawn from a limb.
Synonyms
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| Age of donor (at collection) | 55-59 |
| Collected in | 2017 |
| Source cell line vendor | FSBI Federal Neurosurgical Center |
| Passage number reprogrammed | 1 |
Reprogramming method |
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| Vector type | Non-integrating |
| Vector | Episomal |
| Genes | |
| Is reprogramming vector detectable? |
No |
| Methods used |
PCR
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| Notes on reprogramming vector detection | PCR detection of episomes in cell lines ICGi034-A, ICGo034-B, ICGi034-C |
| 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 selection of colonies was carried out according to morphological criteria. We selected flat monolayer colonies with tightly packed cells with high nuclear/cytoplasmic ratio. Embryonic stem cell-like clones were picked by a glass microcapillary. |
| Derived under xeno-free conditions |
No |
| Derived under GMP? |
No |
| Available as clinical grade? |
No |
Culture Conditions
| Surface coating | Gelatin | ||||||||||||||||||
| Feeder cells |
Mouse embryonic fibroblasts Cellfinder Ont Id: EFO_0004040 |
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| Passage method |
Enzymatically
TrypLE
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| O2 Concentration | 20 % | ||||||||||||||||||
| CO2 Concentration | 5 % | ||||||||||||||||||
| Medium |
Other medium:
Base medium: Knock-out DMEM
Main protein source: Knock-out serum replacement Serum concentration: 15 % 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 |
| Alkaline Phosphatase |
Yes |
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| SSEA-4 |
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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| NANOG |
Yes |
Morphology pictures
Morphol PD30-5-16.tif
Morphology of ICGi034-B iPSC at passage 6
Differentiation Potency
In vitro spontaneous differentiation
Morphology
4 Sp dif PD30-5-16 20p FOXA2_r AFP_g.tif
Immunofluorescent analysis for endoderm markers AFP (green signal) and FOXA2 (red signal) in ICGi034-B iPSC at passage 20. DAPI (blue signal)
Protocol or reference
In vitro spontaneous differentiation
| Marker | Expressed |
| Actin, alpha 2, smooth muscle, aorta |
Yes |
Morphology
2 Sp dif PD30-5-16 20p aSMA_r.tif
Immunofluorescent analysis for mesoderm marker alfa SMA (red signal) in ICGi034-B iPSC at passage 20. DAPI (blue signal)
In vitro spontaneous differentiation
Morphology
4 Sp dif PD30-5-16 20p bIII_r MAP2_g.tif
Immunofluorescent analysis for ectoderm markers TUBB3 (red signel) and MAP2 (green signal) in ICGi034-B iPSC at passage 20. DAPI (blue signal)
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
Karyotyping and G-banding show ICGi034-B PSCs have a normal 46, XX karyotype at passage 8.
Passage number: 8
Karyotyping method:
G-Banding
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Other Genotyping (Cell Line) |
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