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Neuronal Progenitor Cells Derived from XCL-1 MAP2p-Nanoluc-Halotag

ACS-5007

Product category
Human cells
Product type
Reporter-labeled cell
Cell model
Organism
Homo sapiens
Cell type
neural progenitor cell
Morphology
short spindle shape
Tissue
Umbilical cord; CD34+ cord blood
Disease
Normal
Applications
Assay development
Drug development
High-throughput screening
Neuroscience
Stem cell research
Toxicology
Product format
Frozen
Storage conditions
Vapor phase of liquid nitrogen
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Documentation

Biosafety Icon BSL 1

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.

Cells contain Sendai virus (SeV) DNA sequences

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.

Growth Kit for Neural Progenitor Cell Expansion

ACS-3003

Price: $329.00 ea

DMEM: F-12 Medium

30-2006

Price: $24.00 ea

CellMatrix Basement Membrane Gel

ACS-3035

Price: $221.00 ea

Stem Cell Freezing Media

ACS-3020

Price: $55.00 ea

Stem Cell Dissociation Reagent

ACS-3010

Price: $98.00 ea

Detailed product information

General

Specific applications
Neuronal differentiation and drug screening; Quantitative measurement of neuronal differentiation with Nano Luciferase expression under the control of the MAP2 gene promoter.

Characteristics

Cells per vial
≥ 1.0 x 106
Volume
1.0 mL
Growth properties
Adherent
Derivation

ACS-5007 NPCs are derived from NIH NCRM-1 iPSC line

Age
neonate
Ethnicity
White
Gender
Male
Antigen expression
Nestin, Pax-6

Handling information

Unpacking and storage instructions
  1. Check all containers for leakage or breakage.
  2. Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below ­-130°C, preferably in liquid nitrogen vapor, until ready for use.
Complete medium
Complete growth media for Neural Progenitor Cells (NPCs) includes DMEM: F12 (ATCC® 30-2006) supplemented with the Growth Kit for Neural Progenitor Cell Expansion (ATCC® ACS-3003). To make complete NPC medium add the following components of the kit to 464 mL DMEM: F12:

5 mL L-Alanyl-L-Glutamine
5 mL Non-Essential Amino Acids
10 mL NPC Growth Kit Component A
5 mL NPC Growth Kit Component B
1 mL NPC Growth Kit Component C
10 mL NPC Growth Kit Component D
Seeding density
Post-thaw: 8.0 x 104 viable cells/cm2 on CellMatrix-coated dishes/
Subculture: 4.0 x 104 viable cells/cm2 on CellMatrix-coated dishes/plates
Handling procedure

Note: Coat plates with CellMatrix (ATCC® ACS-3035) and culture the NPCs with NPC Growth Medium (ATCC® ACS-3003).

To insure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If, upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at –80°C. Storage at –80°C will result in loss of viability. Preparation of CellMatrix Basement Membrane Gel (ATCC® ACS-3035™) coated plates:
  1. Thaw CellMatrix Basement Membrane Gel on ice or at 4°C
  2. Prepare a 150 µg/ mL working concentration of CellMatrix in cold DMEM: F-12 medium
  3. Add enough CellMatrix solution to cover the surface of the plate (e.g. 1 mL diluted CellMatrix/well of a 12-well plate)
  4. Incubate for 1 hour at 37°C prior to use
Initiation of Cultures
  1. Prepare complete NPC growth medium (ATCC® ACS-3003™) following the instructions in the package and pre-warm that medium as well as DMEM:F12 in a 37°C water bath for 15-30 min. If using a small volume of medium (50 mL or less), warm only the volume needed in a sterile conical tube. Avoid warming complete medium multiple times.
  2. Obtain a 12-well plate with CellMatrix Basement Membrane. Aspirate the CellMatrix medium and directly add 1.5 mL of the complete NPC Growth Medium per well. Place the plate in the incubator for 15 minutes to allow the medium to reach its normal pH (7.0-7.6). Four to five wells of a 6-well plate may be needed for each vial of cells thawed.
  3. Transfer 9 mL of pre-warmed DMEM:F12 into a 15 mL conical tube for recovery of the NPCs from the frozen stock.
  4. Remove cryovial of frozen cells from liquid nitrogen storage.
  5. Thaw the cells by gently swirling in a 37°C water bath. To reduce the possibility of contamination, keep the cap out of the water. Thawing should be rapid (approximately 1 to 2 minutes). Remove the cryovial from water bath when only a few ice crystals are remaining.
  6. Sterilize the cryovial with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions.
  7. Remove cells from the vial using a P1000 micropipette and transfer cells drop-wise into the 15 mL conical tube containing 9 mL DMEM:F12.
  8. Centrifuge cells at 270 x g for 5 minutes at room temperature.
  9. Aspirate the supernatant and discard. Gently tap the bottom of the tube to loosen the cell pellet.
  10. Add 4 mL of the complete NPC Growth Medium to the tube. Gently resuspend the pellet by pipetting up and down 3 or 4 times to make a single-cell suspension.
  11. Perform cell count by a Vi-Cell Analyzer or hemocytometer. Note: Don’t perform cell count by a Vi-Cell Analyzer without removal of serum-free freezing medium.
  12. Seed NPCs at a seeding density of 80,000 viable cells/cm2 (e.g. 0.30 x106/well of a 12-well plate) onto a CellMatrix-coated plate described above.
  13. Incubate the plate at 37°C with 5% CO2 overnight.
  14. Change medium at 100% media change rate (1 mL media/well) next day and every other day thereafter.
  15. Monitor cell growth and passage the cells when they reach ~95% confluency
Note: Don’t passage NPCs when the cells are <85% confluency.
Subculturing procedure

Post thaw day 1, perform a 100% medium change and remove all cells that did not attach. Perform a 100% medium change every other day thereafter. Passage the cells cells with diluted Accutase (50% Accutase and 50% DPBS) when they reach ~95% confluence and reseed the NPCs at 40,000 viable cells/cm2 on CellMatrix-coated dishes/plates.

Cryopreservation

Post thaw day 1, perform a 100% medium change and remove all cells that did not attach. Perform a 100% medium change every other day thereafter. Passage the cells cells with diluted Accutase (50% Accutase and 50% DPBS) when they reach ~95% confluence and reseed the NPCs at 40,000 viable cells/cm2 on CellMatrix-coated dishes/plates.

Quality control specifications

Bacterial and fungal testing
Not detected
Virus testing
Cytomegalovirus (CMV): Not detected
Human papillomavirus (HPV): Not detected
Human immunodeficiency virus 1 (HIV-1): Not detected
Epstein-Barr virus (EBV): Not detected
Hepatitis B virus (HBV): Not detected
Functional tests
Exhibit differentiation potential of NPCs into Tuj1+ early neurons and TH+ dopaminergic neurons
Population doubling capacity
≥15 PDLs
STR profiling
Amelogenin: X,Y
CSF1PO: 9,10
D13S17: 9,10
D16S539: 9,13
D5S818: 14,16
D7S820: 8,9
TH01: 9.3
TPOX: 8,11
Viability
≥ 80%

History

Depositors
ATCC
Year of origin
2015

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.
Astrocyte, oligodendrocyte, and neuron differentiation; drug screening; Quantitative measurement of ealy neuron differentiation.
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

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. HaloTag 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. NanoLuc Label License 

For information on obtaining additional rights, please contact:

ATCC Licensing
Email: [email protected]

MORE INFORMATION ABOUT PERMITS AND RESTRICTIONS

References

Curated Citations

Malik N, et al. Compounds with species and cell type specific toxicity identified in a 2000 compound drug screen of neural stem cells and rat mixed cortical neurons. Neurotoxicology 45: 192-200, 2014. PubMed: 25454721

Frequently Asked Questions

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Telephone

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