ORIONi002-A

iALS-2

General

Cell Line

hPSCreg name ORIONi002-A
Cite as:
ORIONi002-A (RRID:CVCL_C1QB)
Alternative name(s)
iALS-2
Cell line type Human induced pluripotent stem cell (hiPSC)
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Last update 26th July 2022
Notes iALS-2 is a iPSc cell line generated from patient with sporadic form of ALS with a non-viral, polycistronic, synthetic RNA technology.
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Provider

Generator Biomedical center Martin, Jessenius Faculty of Medicine in Martin, COMENIUS UNIVERSITY IN BRATISLAVA (ORION)
Owner Biomedical center Martin, Jessenius Faculty of Medicine in Martin, COMENIUS UNIVERSITY IN BRATISLAVA (ORION)
Distributors
Derivation country Slovakia

External Databases

BioSamples SAMEA12999351
Cellosaurus CVCL_C1QB
Wikidata Q114312638

General Information

Publications
* Is the cell line readily obtainable for third parties?
Yes
Cell line can only be used in: iALS-2 cell line is allowed for research use.
Research use: allowed
Clinical use: not allowed
Commercial use: not allowed

Donor Information

General Donor Information

Sex male
Age of donor (at collection) 45-49
Ethnicity Caucasian

Phenotype and Disease related information (Donor)

Diseases A disease was diagnosed.
sporadic form of amyotrophic lateral sclerosis
The donor is affected.
Synonyms
  • sporadic amyotrophic lateral sclerosis
Family history no family history of ALS
Is the medical history available upon request? yes, it is available upon request
Is clinical information available? yes, it is available

Karyotyping (Donor)

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

Other Genotyping (Donor)

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

External Databases (Donor)

BioSamples SAMEA12999352

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? 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 expressly prevent the derivation of pluripotent stem cells? No
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

Does consent permit research by

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? 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? No
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 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: Medical records will be accessible via medical doctor that was in charge for treating the patient
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: Doctor in charge - Vladimir Nosal, MD, PhD, Martin University Hospital, Slovakia
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? Ethic committee of Jessenius Faculty of Medicine in Martin, Comenius Univeristy in Bratislava, Slovak Republic
Approval number Approval number: EK 1856/2016
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? Ethic committee of Jessenius Faculty of Medicine in Martin, Comenius Univeristy in Bratislava, Slovak Republic
Approval number Approval number: EK 1856/2016
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? MERCK MILLIPORE https://www.merckmillipore.com/SK/sk/product/Simplicon-RNA-Reprogramming-Kit-OKSG,MM_NF-SCR550?ReferrerURL=https%3A%2F%2Fwww.google.com%2F&bd=1
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
Source cell origin
A zone of skin that is part of a lower leg [Automatically generated definition].
Synonyms
  • hind limb middle limb segment skin
  • hind limb zeugopod skin
  • hindlimb zeugopod skin
Source cell type (free text) primary skin fibroblasts isolated with explant culture from skin (lower leg)
Age of donor (at collection) 45-49
Collected in 2018
Passage number reprogrammed P7

Reprogramming method

Vector type Non-integrating
Vector Simplicon™ RNA Reprogramming, single, polycistronic, synthetic, self-replicating RNA strand (contains OKS-iG; Oct4, Klf4, Sox2 and Glis1)
Genes
Is reprogramming vector detectable?
Unknown
Notes on reprogramming vector detection Notes from datasheet: The Simplicon RNA replicon is a synthetic in vitro transcribed RNA expressing all four reprogramming factors (OKS-iG; Oct4, Klf4, Sox2 and Glis1) in a polycistronic transcript that is able to self-replicate for a limited number of cell divisions.

Vector free reprogramming

Type of used vector free reprogramming factor(s)
None

Other

Selection criteria for clones Criteria: morphology of iPSc colonies and expression of stem cell markers
Derived under xeno-free conditions
No
Derived under GMP?
No
Available as clinical grade?
No

Culture Conditions

Surface coating iPSc derivation started with MEFs (mouse embryonic fibroblasts) feeder layer, then cells adapted to feeder-free conditions - Matrigel coating
Feeder cells CF-1 MEFs, EmbryoMax® Primary Mouse Embryonic Fibroblast, PMEF, Strain CF1, Mitomycin C Treated, Passage 3
Cellfinder Ont Id: EFO_0004040
Passage method Mechanically
O2 Concentration 20 %
CO2 Concentration 5 %
Medium mTeSR™ 1
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 Flow Cytometry Enzymatic Assay Expression Profiles
NANOG
Yes
SOX2
Yes
POU5F1 (OCT-4)
Yes
c Myc
Yes
Lin-28
Yes
SSEA-4
Yes
Alkaline Phosphatase
Yes
Score:
Marker Present Absent
mCpG
OCT4
Differentiation Potency
Endoderm
Ont Id: UBERON_0000925
In vitro directed differentiation
Marker Expressed
alpha-fetoprotein
Yes
Morphology
iALS-2 AFP endoderm marker.tif
Expression of alpha-fetoprotein in differentiated iALS-2 cells
Cardiac Muscle Cell
Ont Id: CL_0000746
In vitro directed differentiation
Marker Expressed
alpha smooth muscle actin
Yes
Morphology
iALS-2 alpha-SMA marker for mesoderm.tif
Expression of alpha-SMA in differentiated iALS-2 cells
Ectoderm
Ont Id: UBERON_0000924
In vitro directed differentiation
Marker Expressed
Class III β-tubulin
Unknown
Morphology
iALS-2 TUJ marker for ectoderm.tif
Expression of TUJ in differentiated iALS-2 cells

Microbiology / Virus Screening

Mycoplasma Negative

Genotyping

Karyotyping (Cell Line)

Has the cell line karyotype been analysed?
Yes
46, XY
Passage number: P5
Karyotyping method: G-Banding

Other Genotyping (Cell Line)