iDFF24

General#

Cell Line

hPSCreg Name RCSIi005-A
Alternative name(s)
iDFF24
Cell line type Human induced pluripotent stem cell (hiPSC)
Last update 11th October 2018
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Provider

Generator Royal College of Surgeons in Ireland (RCSI)

External Databases

BioSamples SAMEA5126966
Cellosaurus CVCL_VR61

General Information

Publications
Is the cell line available in principle? Available with restrictions: Available for non-commercial research

Donor Information#

General Donor Information

Sex male

Phenotype and Disease related information (Donor)

Diseases A disease was diagnosed.
diabetes mellitus (primary disease)
Cells collected from non-ulcerated foot region
Synonyms
  • Diabetes mellitus (disorder)
  • Diabetes
  • Diabetes mellitus, NOS
  • DM - Diabetes mellitus
  • Diabetes NOS
show more synonyms

Karyotyping (Donor)

Has the donor karyotype been analysed?
Unkown

Other Genotyping (Donor)

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

External Databases (Donor)

BioSamples SAMEA5126967

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: Jonathan Garlick
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 Jonathan Garlick
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
Alternatives to consent
Alternative consent approval number
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 pertain to a specific research project? No
Details on restriction to research project
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? No
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
Policy for access to genetic information 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? Yes
Please describe how access is provided: Access was provided to Study Research Staff
Does consent permit access to any other source of information about the clinical treatment or health of the donor? No
Contact data, institution, or website:
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? Beth Israel Deaconess Medical Center
Approval number 2011P000226
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
Name of accrediting authority involved?
Approval number
Do you have obligations to third parties in regard to the use of the cell line? Yes
Please describe: Material Transfer agreement in place - cells originated in Tufts University
Are you aware of any further constraints on the use of the donated embryo/tissue or derived cells? No
Further constraints on use
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
Constraints for use or distribution

hIPSC Derivation#

General

Source cell type
fibroblast of dermis
Source cell type (free text) Collected from ulcer free diabetic feet
Passage number reprogrammed 5

Reprogramming method

Vector type Non-integrating
Vector Sendai virus
Genes
Is reprogramming vector detectable?
No
Methods used
RT-PCR
Files and images showing reprogramming vector expressed or silenced

Vector free reprogramming

Type of used vector free reprogramming factor(s)
None

Other

Selection criteria for clones To characterize colonies generated from SeV-infected skin fibroblasts, we picked an average of 20–30 colonies from each cell line. We utilized a live staining approach (Lowry et al., 2008) to select only the colonies that stained positively for both SSEA-4 (Millipore) and TRA-1-81 (BD Biosciences) antibodies confirming that the iPSCs were fully reprogrammed.
Derived under xeno-free conditions
No
Derived under GMP?
No
Available as clinical grade?
No

Culture Conditions#

Surface coating Non coated
Feeder cells gamma-irradiated mouse embryonic fibroblast
Passage method Mechanically
CO2 Concentration 5 %
Medium Other medium:
Base medium: DMEM/F12
Main protein source: Knock-out serum replacement
Serum concentration: 20 %
Supplements
Glutamax 1 %
b-mercaptoethanol 0.1 mM
NEAA 1 %
FGF-2 8 ng/ml
Has Rock inhibitor (Y27632) been used at passage previously with this cell line?
Unkown
Has Rock inhibitor (Y27632) been used at cryo previously with this cell line?
Unkown
Has Rock inhibitor (Y27632) been used at thaw previously with this cell line?
Unkown

Characterisation#

Analysis of Undifferentiated Cells
Marker Expressed Immunostaining RT-PCR FACS Enzymatic Assay Expression Profiles
SSEA-4
Yes
TRA 1-81
Yes
Oct4
Yes
Marker Present Absent
mCpG X
OCT4 X
Morphology pictures
Embryoid body formation; in vivo teratoma
Differentiation Potency
Endoderm
Ont Id: UBERON_0000925
In vivo teratoma
In vitro spontaneous differentiation
Marker Expressed
a-fetoprotein
Yes
Mesoderm
Ont Id: UBERON_0000926
In vivo teratoma
In vitro spontaneous differentiation
Marker Expressed
Vimentin
Yes
Ectoderm
Ont Id: UBERON_0000924
In vivo teratoma
In vitro spontaneous differentiation
Marker Expressed
BIII-tubulin
Yes

Genotyping#

Karyotyping (Cell Line)

Has the cell line karyotype been analysed?
Yes
46XY
Karyotyping method: G-Banding

Other Genotyping (Cell Line)