biological source
human
reprogramming method
retrovirus
description
age (N/A)
manufacturer/tradename
EBiSC™
gender
female
growth mode
adherent (pluripotent)
technique(s)
cell culture | stem cell: suitable
relevant disease(s)
Parkinson′s disease
shipped in
dry ice
storage temp.
−196°C
General description
Induced pluripotent stem cells (iPSCs) are adult cells that have been reprogrammed to an embryonic stem cell–like state. The cells can replicate indefinitely or, under controlled conditions, can be differentiated into any other cell type such as nerve, heart or liver cells. Medical researchers are able to use iPS cells to test how different patients might respond to new drugs or to analyse how genetic diseases develop.
The EBiSC stem cell bank is a collection of human iPS cells available to academic and commercial researchers for use in disease modelling and other forms of stem cell research. The initial collection has been generated from a wide range of donors representing specific disease backgrounds and healthy controls. EBiSC has established many routine procedures for collecting, expanding and characterizing human iPS cell lines. The stem cell bank includes iPSC cell lines derived from neurodegenerative diseases (Alzheimer′s Disease, Parkinson′s Disease, Dementia, Motor Neuron Disease (ALS) - and Huntington′s Disease), eye and heart diseases, and lines from healthy control donors for age and sex matching.
The EBiSC stem cell bank is a collection of human iPS cells available to academic and commercial researchers for use in disease modelling and other forms of stem cell research. The initial collection has been generated from a wide range of donors representing specific disease backgrounds and healthy controls. EBiSC has established many routine procedures for collecting, expanding and characterizing human iPS cell lines. The stem cell bank includes iPSC cell lines derived from neurodegenerative diseases (Alzheimer′s Disease, Parkinson′s Disease, Dementia, Motor Neuron Disease (ALS) - and Huntington′s Disease), eye and heart diseases, and lines from healthy control donors for age and sex matching.
Cell Line Origin
Depositor
University of Edinburgh
University of Edinburgh
Cell Line Description
Derivation
Primary cell type: -
Reprogramming method
Vector type: Integrating
Vector: Virus
Virus type: Retrovirus
Gene list:
KLF4
MYC
POU5F1
SOX2
Have the reprogramming vectors been silenced: Yes
Merthods used: rtpcr
Xeno free conditions: no
Derived under gmp: no
Available as clinical grade: no
Characterization
Microbiology / Virus Screening
HIV 1: Not done
HIV 2: Not done
Hepatitis B: Not done
Hepatitis C: Not done
Mycoplasma: Negative
Sterility
Inoculation for microbiological growth: No Contaminants Detected
Mycoplasma: Not Detected
Viability: Viable post-cryopreservation
Genotyping
STR/Fingerprinting: A 16 allele profile has been recorded and data is available upon request, after cell line purchase.
Genetic Modification
Disease/phenotype related modifications
Disease: Parkinson′s disease
Details: This is a PD line, with the control being NAS lines and CRISPR/Cas9-corrected AST22 lines
Type of modification: Gene knock-out
Gene: SNCA
Chromosome location: 4q22.1
Delivery method: CRISPR-associated (CRISPR/Cas) System
Description: CRISPR/Cas was employed to create a mutation in Exon2, disrupting the ATG start site and some sequence 5′ to the coding start of SCNA. PCR amplicons of the CRISPR site were cloned using the TOPO cloning technique. Sequencing of 8 TOPO clones detected 2 deletions and 1 insertion; however, sequencing of the CRISPR sites shows that only one of these modifications disrupts the ATG site. This indicates 1 allele was putatively knocked out, and 3 alleles remain. Additional comment: there are two additional ATG sites located immediately downstream of the normal ATG start site, one of which is in-frame (9 bases) to the initial ATG. It is possible that inactivation of the first ATG start site might not be completely sufficient to prevent translation from a second downstream ATG.
Primary cell type: -
Reprogramming method
Vector type: Integrating
Vector: Virus
Virus type: Retrovirus
Gene list:
KLF4
MYC
POU5F1
SOX2
Have the reprogramming vectors been silenced: Yes
Merthods used: rtpcr
Xeno free conditions: no
Derived under gmp: no
Available as clinical grade: no
Characterization
Microbiology / Virus Screening
HIV 1: Not done
HIV 2: Not done
Hepatitis B: Not done
Hepatitis C: Not done
Mycoplasma: Negative
Sterility
Inoculation for microbiological growth: No Contaminants Detected
Mycoplasma: Not Detected
Viability: Viable post-cryopreservation
Genotyping
STR/Fingerprinting: A 16 allele profile has been recorded and data is available upon request, after cell line purchase.
Genetic Modification
Disease/phenotype related modifications
Disease: Parkinson′s disease
Details: This is a PD line, with the control being NAS lines and CRISPR/Cas9-corrected AST22 lines
Type of modification: Gene knock-out
Gene: SNCA
Chromosome location: 4q22.1
Delivery method: CRISPR-associated (CRISPR/Cas) System
Description: CRISPR/Cas was employed to create a mutation in Exon2, disrupting the ATG start site and some sequence 5′ to the coding start of SCNA. PCR amplicons of the CRISPR site were cloned using the TOPO cloning technique. Sequencing of 8 TOPO clones detected 2 deletions and 1 insertion; however, sequencing of the CRISPR sites shows that only one of these modifications disrupts the ATG site. This indicates 1 allele was putatively knocked out, and 3 alleles remain. Additional comment: there are two additional ATG sites located immediately downstream of the normal ATG start site, one of which is in-frame (9 bases) to the initial ATG. It is possible that inactivation of the first ATG start site might not be completely sufficient to prevent translation from a second downstream ATG.
Linkage
Note: EAUA and CLIP must be completed before order fulfillment
Subculture Routine
Medium: mTeSR®
Passage method: EDTA
Matrix: Matrigel® / Geltrex®
CO2 concentration: 5%
O2 concentration: 20%
Temperature: 37°C
Passage method: EDTA
Matrix: Matrigel® / Geltrex®
CO2 concentration: 5%
O2 concentration: 20%
Temperature: 37°C
Legal Information
EBiSC is a trademark of Fraunhofer-Gesellschaft
GELTREX is a registered trademark of Life Technologies Corporation
Matrigel is a registered trademark of Corning, Inc.
mTeSR is a registered trademark of WiCell Research Institute, Inc.
Storage Class Code
10 - Combustible liquids
WGK
WGK 3
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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