HLHS_375

General#

Cell Line

hPSCreg Name
MRIi018-A
Alternative name(s)
HLHS_375
Cell line type Human induced pluripotent stem cell (hiPSC)
Last update 19th May 2021
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Provider

Generator Klinikum rechts der Isar (MRI)
Derivation country Germany

External Databases

BioSamples SAMEA8944460
CLO CLO_0103312

General Information

* Is the cell line readily obtainable for third parties?
No

Donor Information#

General Donor Information

Sex male
Ethnicity Caucasian

Phenotype and Disease related information (Donor)

Diseases A disease was diagnosed.
Hypoplastic left heart syndrome (primary disease)
The donor is a carrier of a disease-associated mutation and affected.
Synonyms
  • HLHS
Disease associated phenotypes
  • The disease-associated phenotype of HLHS includes a hypoplastic left ventricle, a hypoplastic aorta, as well as impairment of the mitral and aortic valve. No ventricular septal defect has been detected.
Family history No abnormalities have been described regarding the family history.
Is the medical history available upon request? Clinical information is available upon request.
Is clinical information available? Clinical information is available upon request.

Karyotyping (Donor)

Has the donor karyotype been analysed?
No

Other Genotyping (Donor)

Is there genome-wide genotyping or functional data available?
Yes
Exome sequencing
Three de novo mutations found. BAI2: c.800G>A; DENND5B: c.2396A>G; SYBU: c.278C>A.

External Databases (Donor)

BioSamples SAMEA8944459

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? Yes
Details on restriction to research project
Does consent permit unforeseen future research, without further consent? 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? 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? No
Does consent expressly permit storage of cells derived from the donated embryo/tissue for an unlimited time? 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? No
a public organisation? No
a non-profit company? No
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? No
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? 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: German Heart Center Munich, Department of Cardiovascular Surgery, Institute Insure
Does consent permit access to any other source of information about the clinical treatment or health of the donor? Yes
Contact data, institution, or website: German Heart Center Munich, Department of Cardiovascular Surgery, Institute Insure
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? German Heart Center Munich, Department of Cardiovascular Surgery, Institute Insure
Approval number 5943/13, 2919/10
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? German Heart Center Munich, Department of Cardiovascular Surgery, Institute Insure
Approval number 5943/13, 2919/10
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? Thermofisher
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 type
skin fibroblast

Reprogramming method

Vector type Non-integrating
Vector Sendai virus
Is reprogramming vector detectable?
Yes
Methods used
RT-PCR
Files and images showing reprogramming vector expressed or silenced

Vector free reprogramming

Other

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

Culture Conditions#

Surface coating Matrigel/Geltrex
Feeder cells
No
Passage method Enzyme-free cell dissociation
EDTA
CO2 Concentration 5 %
Medium Essential 8™
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?
Yes
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
TRA 1-81
Yes
SOX2
Yes
NANOG
Yes
KLF4 – Kruppel like factor 4
Yes
REX1 - RNA exonuclease 1 homolog
Yes
OCT4 - POU class 5 homeobox 1
Yes
c-MYC - MYC proto-oncogene, bHLH transcription factor
Yes
Score:
Marker Present Absent
mCpG
OCT4
Morphology pictures
Representative colony of the HLHS_375 iPSC line.
Differentiation Potency
Endoderm
Ont Id: UBERON_0000925
In vitro spontaneous differentiation
Protocol or reference
AFP_expression.png
Quantitative RT-PCR for the AFP gene expression representative of the endoderm germ layer upon spontaneous embryoid-body differentiation (D21 vs D0).
SOX7_expression.png
Quantitative RT-PCR for the SOX7 gene expression representative of the endoderm germ layer upon spontaneous embryoid-body differentiation (D21 vs D0).
Mesoderm
Ont Id: UBERON_0000926
In vitro spontaneous differentiation
Protocol or reference
ACTA2_expression.png
Quantitative RT-PCR for the ACTA2 gene expression representative of the mesoderm germ layer upon spontaneous embryoid-body differentiation (D21 vs D0).
CD31_expression.png
Quantitative RT-PCR for the CD31 (PECAM) gene expression representative of the mesoderm germ layer upon spontaneous embryoid-body differentiation (D21 vs D0).
Ectoderm
Ont Id: UBERON_0000924
In vitro spontaneous differentiation
Marker Expressed
KRT14 keratin 14
Yes
TH tyrosine hydroxylase
Yes
Protocol or reference
KRT14_expression.png
Quantitative RT-PCR for the KTR14 gene expression representative of the ectoderm germ layer upon spontaneous embryoid-body differentiation (D21 vs D0).
TH_expression.png
Quantitative RT-PCR for the TH gene expression representative of the ectoderm germ layer upon spontaneous embryoid-body differentiation (D21 vs D0).

Microbiology / Virus Screening

HIV 1 Negative
Hepatitis B Negative
Hepatitis C Negative
Mycoplasma Negative

Genotyping#

Karyotyping (Cell Line)

Has the cell line karyotype been analysed?
Yes

Other Genotyping (Cell Line)