iPSC_FB_M45-50_C3

General#

Cell Line

hPSCreg Name
BBANTWi001-C
Alternative name(s)
iPSC_FB_M45-50_C3
Cell line type Human induced pluripotent stem cell (hiPSC)
Last update 25th June 2021
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Provider

Generator Biobank Antwerpen (BBANTW)
Owner Center of Medical Genetics Antwerp (CMGANT)

External Databases

Cellosaurus CVCL_ZX27
BioSamples SAMEA9269364
CLO CLO_0103349

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 male
Ethnicity white Belgian

Phenotype and Disease related information (Donor)

Diseases No disease was diagnosed.

Karyotyping (Donor)

Has the donor karyotype been analysed?
Yes
46, XY. No clinically significant abnormalities observed.
Karyotyping method: Molecular karyotyping by SNP array
http://

Other Genotyping (Donor)

Is there genome-wide genotyping or functional data available?
Yes
SNP typing array
Skin fibroblast DNA sample was genotyped with Illumina's HumanCytoSNP-12 v2.1. No clinically significant CNVs observed.

Donor Relations

Other cell lines of this donor

External Databases (Donor)

BioSamples SAMEA9269223

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
Confirm that consent was obtained by a qualified professional Yes
Has the donor agreed to be re-contacted? Yes
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? Yes
Details on restriction to research project Study of inherited cardiovascular diseases
Does consent permit unforeseen future research, without further consent? 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 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? No
Has the donor consented to receive information discovered during use of donated embryo/tissue that has significant health implications for the donor? 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? Yes
Please describe how access is provided: Through the treating physician
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? UZA ethics committee
Approval number EC 18/12/166
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? UZA ethics committee
Approval number EC 18/12/166
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? CytoTune™-iPS 2.0 Sendai Reprogramming Kit from Invitrogen / ThermoFisher Scientific

hIPSC Derivation#

General

Source cell type
skin fibroblast
Source cell origin
upper arm
Source cell type (free text) Skin biopsy was taken on inner side of upper arm
Passage number reprogrammed 6

Reprogramming method

Vector type Non-integrating
Vector Sendai virus
Is reprogramming vector detectable?
No
Methods used
RT-PCR
Notes on reprogramming vector detection SeV not detected on passage 12

Vector free reprogramming

Other

Selection criteria for clones Clones were picked based on morphology (round, flat colonies with smooth edges and tightly packed cells) for 5 rounds and subsequently passaged using EDTA for 5 more rounds. Only clones with nice morphology and no differentiation after these 10 passaging rounds were selected.
Derived under xeno-free conditions
No
Derived under GMP?
Unknown
Available as clinical grade?
No

Culture Conditions#

Surface coating Matrigel/Geltrex
Feeder cells
No
Passage method Enzyme-free cell dissociation
EDTA
O2 Concentration 21 %
CO2 Concentration 5 %
Medium Essential 8™ Flex
Has Rock inhibitor (Y27632) been used at passage previously with this cell line?
Yes
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?
Yes

Characterisation#

Analysis of Undifferentiated Cells
Marker Expressed Immunostaining RT-PCR FACS Enzymatic Assay Expression Profiles
POU5F1 (OCT-4)
Yes
TRA 1-60
Yes
TRA 1-81
Yes
NANOG
Yes
SOX2
Yes
Morphology pictures
Brieghtfield image of 60% confluent iPSC culture
Differentiation Potency
Endoderm
Ont Id: UBERON_0000925
In vitro spontaneous differentiation
Marker Expressed
SOX17
Yes
GATA6
Yes
CXCR4
Yes
Mesoderm
Ont Id: UBERON_0000926
In vitro spontaneous differentiation
Marker Expressed
DCN
Yes
VIM
Yes
PECAM1
Yes
Cardiac Muscle Cell
Ont Id: CL_0000746
In vitro directed differentiation
Marker Expressed
alpha-actinin 2
Yes
Troponin T, cardiac type
Yes
SCN5A
Unknown
NKX2-5
Unknown
MYL2
Unknown
Ectoderm
Ont Id: UBERON_0000924
In vitro spontaneous differentiation
Marker Expressed
PAX6
Yes
HES5
Yes
NEUROD1
Yes

Microbiology / Virus Screening

Mycoplasma Negative

Genotyping#

Karyotyping (Cell Line)

Has the cell line karyotype been analysed?
Yes
46, XY. No clinically significant abnormalities observed.
Passage number: 12
Karyotyping method: Molecular karyotyping by SNP array
http://

Other Genotyping (Cell Line)

Is there genome-wide genotyping or functional data available?
Yes
SNP typing array
Genotyped with Illumina's HumanCytoSNP-12 v2.1. No clinically significant CNVs observed.