RCPCMi001-A

ATXN1ACL4

The cell line is not validated yet.

General#

Cell Line

hPSCreg Name RCPCMi001-A
Alternative name(s)
ATXN1ACL4
Cell line type Human induced pluripotent stem cell (hiPSC)
Last update 22nd May 2019
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Provider

Generator Federal Research and Clinical Center of Physical-Chemical Medicine (RCPCM)

External Databases

BioSamples SAMEA5176975
Cellosaurus CVCL_UK94

General Information

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

Donor Information#

General Donor Information

Sex female
Ethnicity Caucasian

Phenotype and Disease related information (Donor)

Diseases A disease was diagnosed.
Ataxia
Spinocerebellar ataxia type 1 (SCA-1) belongs to the class of polyglutamine neurodegenerative diseases. This is an autosomal dominant disease caused by the expansion of CAG repeats in the ATXN1 gene encoding the polyglutamine tract in ataxin-1 protein (Orr et al., 1993). The exact functions of both normal and mutant ATXN1 are largely unknown. SCA-1 is caused by Purkinje cell death and degradation of the olive-cerebellar tracts, which leads to impaired coordination of movements, and cognitive disorders. Usually the onset of the disease occurs in 30-40 years old. The severity of the disease depends on the length of the CAG tract (Durr, 2010).
The donor is a carrier of a disease-associated mutation and affected.
Synonyms
  • ataxia
  • Ataxia

External Databases (Donor)

BioSamples SAMEA5176974

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: Research Center of Neurology, Deputy Head of Institute S.N Illarioshkin +7 (495) 374-77-76
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 Research Center of Neurology, Deputy Head of Institute S.N Illarioshkin +7 (495) 374-77-76
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
Alternative consent approval number
Please indicate whether the data associated with the donated material has been pseudonymised or anonymised. anonymised
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 permit storage of 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? No
an academic institution? Yes
a public organisation? Yes
a non-profit company? Yes
Does consent expressly permit collection of genetic information? Yes
Does consent expressly permit storage of genetic information? Yes
Policy for access to genetic information Open Access
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? Federal Scientific Center of Neurology
Approval number na
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
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 line name Skin fibroblasts
Source cell type
fibroblast of dermis
Source cell origin
dermis
The dermis is a layer of skin between the epidermis (with which it makes up the skin) and subcutaneous tissues, and is composed of two layers, the papillary and reticular dermis[WP].
Synonyms
  • vertebrate dermis
Source cell type (free text) iPSC lines from skin fibroblasts collected from two SCA-1 female patients
Passage number reprogrammed 3

Reprogramming method

Vector type Integrating
Vector Virus (Lentivirus)
Genes
Is the used vector excisable?
Yes
Absence of reprogramming vector(s)?
Unknown
Reprogramming vectors silenced?
Methods used
Immunostaining, RT-PCR
Files and images showing reprogramming vector expressed or silenced

Vector free reprogramming

Other

Selection criteria for clones At passage 3 fibroblasts were transduced with lentiviral vectors LeGO-hOCT4, LeGO-hSOX2, LeGO-hc-Myc, LeGO-hKLF4 as described in (Chestkov et al., 2014). During 4 days after transduction valproic acid (Stemgent) was added in the culture medium to the concentration 1 mM. At day 5 cells were replated 1: 6, and cultivated on mitomycin C (Sigma) treated mouse embryonic fibroblasts. The next day after replating culture medium was changed to DMEM/F12 (PanEco), 20% KO Serum replacement (Invitrogen, USA), 0.1mM β-mercaptoethanol (Sigma), 1% NEAA (Hyclone), bFGF 4 ng/ml (PeproTech), 50 units/mL penicillin, and 50 mkg/mL streptomycin (PanEco). Cells were cultured for 12-15 days. Individual colonies were mechanically picked and cultured in TeSRE8 (StemCell Technologies) on Matrigel (Corning).
Derived under xeno-free conditions
Yes
Derived under GMP?
No
Available as clinical grade?
No

Culture Conditions#

Surface coating Matrigel/Geltrex
Feeder cells
No
Passage method Enzymatically
TrypLE
CO2 Concentration 5 %
Medium TeSR™ E8™
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 1-60
Yes
Marker Present Absent
mCpG
OCT4 X
Morphology pictures

Microbiology / Virus Screening

HIV 1 Negative
HIV 2 Negative
Hepatitis B Negative
Hepatitis C Negative
Mycoplasma Negative

Genotyping#

Karyotyping (Cell Line)

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

Other Genotyping (Cell Line)