General#

Cell Line

hPSCreg Name IDISi001-A
Cell line type Human induced pluripotent stem cell (hiPSC)
Last update 22nd May 2019
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Provider

Generator Health Research Institute of Santiago de Compostela (IDIS)
Owner Health Research Institute of Santiago de Compostela (IDIS)

External Databases

Cellosaurus CVCL_RV23
BioSamples SAMEA4675695

General Information

Publications
* Is the cell line readily obtainable for third parties?
Yes
Research: allowed
Clinical: allowed
Commercial: allowed

Donor Information#

General Donor Information

Sex male
Ethnicity Caucasian

Phenotype and Disease related information (Donor)

Diseases A disease was diagnosed.
CADASIL
The donor is a carrier of a disease-associated mutation.
Synonyms
  • Hereditary multi-infarct dementia
  • Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy

Karyotyping (Donor)

Has the donor karyotype been analysed?
Unknown

External Databases (Donor)

BioSamples SAMEA4675696

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: Dr. Francisco Campos Pérez (francisco.campos.perez@sergas.es)
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 Dr. Francisco Campos Pérez (francisco.campos.perez@sergas.es)
Alternatives to consent
Alternative consent approval number
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? No
Details on restriction to research project
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 CELLS DERIVED FROM THE DONATED BIOSAMPLE from being made available to researchers anywhere in the world? Yes
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
Please describe how access is provided:
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? Commission on Guarantees for the Donation and Use of Human Cells and Tissues of Instituto de Salud Carlos III
Approval number 428-346-1
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? Commission on Guarantees for the Donation and Use of Human Cells and Tissues of Instituto de Salud Carlos III
Approval number 428-346-1
Do you have obligations to third parties in regard to the use of the cell line? No
Please describe:
Further constraints on use
For generation of the cell line, who was the supplier of any recombined DNA vectors or commercial kits used?
Constraints for use or distribution

hIPSC Derivation#

General

Source cell type
mononuclear cell
A leukocyte with a single non-segmented nucleus in the mature form.
Synonyms
  • mononuclear leukocyte

Reprogramming method

Vector type Non-integrating
Vector Sendai virus
Genes
Is reprogramming vector detectable?
Yes
Methods used
PCR
Notes on reprogramming vector detection Detection of the DNA vector by PCR of a sample of PBMCs 24h after infection. Absence of the vector was checked by PCR of several clones of the new line at passage 5
Files and images showing reprogramming vector expressed or silenced

Vector free reprogramming

Other

Selection criteria for clones For the first and second passages, colonies were selected and pickedwith the help of an Origio Stripper (Origio)with Stripper tips (Origio) of 175 μm in diameter. Later passages were performed mechanically, using cell lifters.
Derived under xeno-free conditions
Unknown
Derived under GMP?
Unknown
Available as clinical grade?
Unknown

Culture Conditions#

Surface coating Matrigel/Geltrex
Feeder cells
No
Passage method Mechanically
CO2 Concentration 5 %
Medium mTeSR™ 1
Supplements
mTeSR1 Supplemental mix 100 ml
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 FACS Enzymatic Assay Expression Profiles
Alkaline Phosphatase
Yes
NANOG
Yes
POU5F1 (OCT-4)
Yes
SOX2
Yes
SSEA-4
Yes
TRA 1-60
Yes
Score:
Marker Present Absent
mCpG
OCT4
Differentiation Potency
Endoderm
Ont Id: UBERON_0000925
In vitro spontaneous differentiation
Marker Expressed
FOXA2
Yes
Morphology
EBs 05-05-17 HF_Image018.tif
FOXA2 (Green) + DAPI (Blue)
Mesoderm
Ont Id: UBERON_0000926
In vitro spontaneous differentiation
Marker Expressed
Vimentin
Unknown
Morphology
EBs 05-05-17 HF_Image032.tif
Vimentin (Green) + DAPI (Blue)
EBs 05-05-17 HF_Image028.tif
Vimentin (Green) + DAPI (Blue)
Ectoderm
Ont Id: UBERON_0000924
In vitro spontaneous differentiation
Marker Expressed
NES
Unknown
Morphology
EBs 19-05-17_Image012.tif
Nestin (Green) + DAPI (Blue)
EBs 19-05-17_Image025.tif
Nestin (Green) + DAPI (Blue)

Microbiology / Virus Screening

Mycoplasma Negative

Genotyping#

Karyotyping (Cell Line)

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

Other Genotyping (Cell Line)