iPDK

General#

Cell Line

hPSCreg Name RCSIi001-A
Alternative name(s)
iPDK
Cell line type Human induced pluripotent stem cell (hiPSC)
Last update 11th June 2020
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Provider

Generator Royal College of Surgeons in Ireland (RCSI)

External Databases

BioSamples SAMEA5100026
Cellosaurus CVCL_YX99

General Information

* Is the cell line readily obtainable for third parties?
Yes
Research: allowed
Clinical: not allowed
Commercial: not allowed

Donor Information#

General Donor Information

Sex unknown

Phenotype and Disease related information (Donor)

Diseases No disease was diagnosed.

Karyotyping (Donor)

Has the donor karyotype been analysed?
Unknown

Other Genotyping (Donor)

Is there genome-wide genotyping or functional data available?
No

External Databases (Donor)

BioSamples SAMEA5100025

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
Provide contact information of the holder of the original Donor Information Sheet: ATCC
Do you (Depositor/Provider) hold a copy of the SIGNED Donor Consent Form? No
If you do not hold the SIGNED Donor Consent Form, do you know who does? Yes
Contact information / weblink https://www.lgcstandards-atcc.org/?geo_country=ie
If you do not hold the SIGNED Donor Consent Form, have you obtained a copy of the unsigned Donor Consent Form from the holder? 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
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. anonymised
Does consent explicitly allow the derivation of pluripotent stem cells? Yes
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
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? No
Does consent expressly permit storage of genetic information? No
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
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? ATTC
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? No
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? Invitrogen
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

Source cell line name BJ
Other names
Source cell type
fibroblast
A connective tissue cell which secretes an extracellular matrix rich in collagen and other macromolecules. Flattened and irregular in outline with branching processes; appear fusiform or spindle-shaped.
Source cell origin
Foreskin
Source cell type (free text) ATCC® CRL-2522
Source cell line vendor ATCC
Passage number reprogrammed p8

Reprogramming method

Vector type Integrating
Vector Virus (Lentivirus)
Genes
Is the used vector excisable?
Yes
Absence of reprogramming vector(s)?
No
Reprogramming vectors silenced?
Yes
Methods used
Immunostaining, PCR
Notes on reprogramming vector silencing doxycycline (Dox)-inducible lentiviral vectors used

Vector free reprogramming

Type of used vector free reprogramming factor(s)
None

Other

Derived under xeno-free conditions
Yes
Derived under GMP?
No
Available as clinical grade?
No

Culture Conditions#

Surface coating Gelatin
Feeder cells Mouse embryonic fibroblast cell
Passage method Mechanically
Medium Other medium:
Base medium: 80% Knockout DMEM
Main protein source: Knock-out serum replacement
Serum concentration: 10 %
Supplements
Plasmanate 8 %
FCS 5 %
Glutamax 2 mM
NEAA 1 %
Pen/Strep 50 µg/ml
.055 beta-mercaptoenthanol mM
recombinant hLIF 12 ng/ml
bFGF 5 ng/ml
Has Rock inhibitor (Y27632) been used at passage previously with this cell line?
Unknown
Has Rock inhibitor (Y27632) been used at cryo previously with this cell line?
Unknown
Has Rock inhibitor (Y27632) been used at thaw previously with this cell line?
Unknown

Characterisation#

Analysis of Undifferentiated Cells
Marker Expressed Immunostaining RT-PCR FACS Enzymatic Assay Expression Profiles
TRA 1-81
Yes
see: https://ac.els-cdn.com/S1934590908004013/1-s2.0-S1934590908004013-main.pdf?_tid=d27d2580-f893-4acd-a59c-ca4b914087d1&acdnat=1537976369_f40f42d19eb20f05f9e32b0a0bf5b747 The NANOG and OCT4 promoters in fibroblastderived hiPSCs were demethylated to a similar extent as in hESCs, in contrast to the highly methylated promoters in BJ fibroblasts (Figure S1B), thus demonstrating epigenetic reprogramming in hiPSCs. Global analysis of gene expression in fibroblast- derived hiPSCs, hESCs, and fibroblasts was conducted by microarray through comparison of differentially expressed genes between fibroblasts and hESCs (Figure 1C), indicating that hiPSCs had repressed the fibroblast program of gene expression and reactivated an embryonic program of transcription.
Transcriptome Characterisation
Differentiation Potency

Genotyping#

Karyotyping (Cell Line)

Has the cell line karyotype been analysed?
Unknown

Other Genotyping (Cell Line)