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SNCA3X_1KO_C2

SNCA3X_1KO_C2

SNCA3X_1KO_C2 is a CRISPR-engineered induced pluripotent stem cell (iPSC) with SNCA frameshift mutations from a donor with an SNCA triplication. This cell line is part of a panel of cells that can be used to study alpha-synuclein gene expression dosage from the endogenous locus and has applications in Parkinson's disease research.
Product category
Human cells
Product type
Cell model
Organism
Homo sapiens, human
Tissue
Skin
Disease
Parkinsons disease
Applications
Neuroscience
Product format
Frozen
Storage conditions
Vapor phase of liquid nitrogen
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Documentation

ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.

ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.

Required Products

These products are vital for the proper use of this item and have been confirmed as effective in supporting functionality. If you use alternative products, the quality and effectiveness of the item may be affected.

Detailed product information

General

Specific applications
Parkinson’s disease research, neurodegenerative disease drug discovery and development. Lewy body disease.

Characteristics

Cells per vial
≥ 1.0 x 106
Growth properties
Adherent
Age
42 years
Ethnicity
White
Gender
Male
Comments
This cell line is part of a CRISPR-engineered a panel of iPSCs with SNCA frameshift mutations from a donor with an SNCA triplication to study alpha-synuclein gene expression from the endogenous locus.

Handling information

Complete medium

The basal medium for this cell line is StemFlex™ Basal Medium. To make the complete medium add the following to 450 mL basal medium:

  • 50 mL StemFlex™ Supplement 10X 
  • 1 µM of Thiazovivin: 1 mM Thiazovivin/DMSO solution - prepared using 1 mg Thiazovivin/ 3.1212 ml DMSO

Note: Thiazovivin/DMSO solution is used only at Start-Up and Day 0 of all subcultures and removed next day - substituted with StemFlex™ Basal Medium.

Temperature
37°C
Atmosphere
95% Air, 5% CO2
Subculturing procedure
Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.  Corning® T-75 flasks (catalog #430641) are recommended for subculturing this product.
    Notes: CBM coated culture vessels used. Do NOT use CellSTACKs Note on dissociation medium
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with PBS.
  3. Add 2.0 to 3.0 mL of 0.5mM EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
    Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. Centrifuge cells to remove dissociation agent and resuspend in complete media .
  6. Add appropriate aliquots of the cell suspension to new CBM coated culture vessels.
    Cultures can be established between 2 x 104 and 1 x 105 viable cells/cm2. Do not exceed 7 x 104 cells/cm2.
  7. Incubate cultures at 37°C.
Interval: Maintain cultures at a cell concentration between 1.5 X 104 and 6 X 104 cell/cm2.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:8 is recommended
Medium Renewal: 2 to 3 times per week
Notes: Prepare in advance CBM coated cell culture flasks prior to subculturing. Media changes must be performed every other day. Subculture at ≤ 70% confluence. For all startups dilute 1 mL of cell vial with 4 mL of complete medium and perform cell counts prior to centrifugation.
Reagents for cryopreservation
BAMBANKER™ Serum Free Cell Freezing Medium (Fisher Scientific catalog # NC9582225)

Quality control specifications

Bacterial and fungal testing
Not detected
Mycoplasma contamination
Not detected
Virus testing
Cytomegalovirus (CMV): Not detected
Epstein-Barr virus (EBV): Not detected
Hepatitis B virus (HBV): Not detected
Human Immunodeficiency virus (HIV): Not detected
Human papillomavirus (HPV): Not detected
Functional tests

Pluritest:

Pluripotency Score > 20, Novelty Score < 1.80. Specification: requires one set of Pluritest testing results only.

Surface markers; Flow Cytometry:

SSEA-1: Specification: Report Results
SSEA-4: Specification: ≥ 85% of cells expressing marker TRA-1-60: Specification: ≥ 85% of cells expressing marker
Negative Control; Specification: Unstained cells

 

Genotyping:

Specification: Pass.

STR profiling
Amelogenin: X,Y
CSF1PO: 10,11
D13S317: 13
D16S539: 11
D5S818: 11
D7S820: 11,12
THO1: 6,9.3
TPOX: 8,11
vWA: 17,18

History

Depositors
Birgitt Schüle, MD
Department of Pathology
Stanford University School of Medicine
300 Pasteur Dr.
Stanford, CA 94305
Year of origin
2019
Special collection
Michael J Fox Foundation (MJFF)

Legal disclaimers

Intended use
This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
Warranty

The product is provided 'AS IS' and the viability of ATCC® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid.  Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.

Disclaimers

This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Any proposed commercial use is prohibited without a license from ATCC.

While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.

This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.

Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at www.atcc.org.

Permits & Restrictions

Material Transfer Agreement Addendum for Michael J Fox Products

For every order of this item, you must provide a signed Material Transfer Agreement Addendum for Michael J Fox Foundation products. We cannot ship this item until we receive this addendum. Email the signed addendum to [email protected] with a reference to both your account and sales order numbers. Once received, your addendum will be reviewed, and this item will be released for shipment if all requirements are met. If you need assistance with your order, please contact our Customer Care team or your applicable distributor.

This material is subject to the following restrictions in addition to those outlined in the ATCC Material Transfer Agreement:

  1. iPS Label License 

For information on obtaining additional rights, please contact:

ATCC Licensing
Email: [email protected]

This material is subject to the following restrictions in addition to those outlined in the ATCC Material Transfer Agreement:

  1. CRISPR Label License 

For information on obtaining additional rights, please contact:

ATCC Licensing
Email: [email protected]

This material is subject to the following restrictions in addition to those outlined in the ATCC Material Transfer Agreement:

  1. CRISPR Label License, ERS Genomics 

For information on obtaining additional rights, please contact:

ATCC Licensing
Email: [email protected]

MORE INFORMATION ABOUT PERMITS AND RESTRICTIONS

Frequently Asked Questions

References

Curated Citations

Zafar F, et al. Isogenic human SNCA gene dosage induced pluripotent stem cells to model Parkinson’s disease. Stem Cell Res (60): 102733, 2022. PubMed: 35263701

For product-related inquiries and issues, contact Technical Service:

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