hTERT NF1 ipnNF95.11c (ATCC® CRL-3391)

Organism: Homo sapiens, human  /  Cell Type: Schwann cell  /  Tissue: Unaffected (non-tumor) peripheral nerve from NF1 patient. Primary site: peripheral nerve in trunk.  /  Disease: plexiform neurofibroma; tumor not malignant

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Organism Homo sapiens, human
Tissue Unaffected (non-tumor) peripheral nerve from NF1 patient. Primary site: peripheral nerve in trunk.
Cell Type Schwann cell
Product Format frozen 1.0 mL
Morphology fibroblast/spindle-like
Culture Properties adherent
Biosafety Level 2

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Disease plexiform neurofibroma; tumor not malignant
Age 32 years old
Gender male
Ethnicity white
Applications

Through generous support from the Neurofibromatosis Therapeutic Acceleration Program (NTAP), purchasers of these cells are eligible to receive up to a 100% reimbursement from NTAP.  For additional information and to apply for reimbursement after a purchase, please contact NTAP at info@n-tap.org

For study of Schwann cell biology, diseases and tumors. These cells are invaluable tools for dissecting the processes contributing to the formation of neurofibromas and possibly other diseases, and the development of disease models and therapeutics.

The collection contains “normal” (wild-type) NF1(+/+),  NF1(+/-) (heterozygous), and NF1(-/-) homozygous (neurofibroma derived) human Schwann cell lines.  (NOTE: Cell-lines generated under auspices of the lab of Dr. Peggy Wallace, University of Florida, with funding support by the Neurofibromatosis Therapeutic Acceleration Program (NTAP) at Johns Hopkins University)

Storage Conditions liquid nitrogen vapor phase
Karyotype 46,XY (p24)
Images
Comments

A team of researchers from the University of Florida, the National Center for Advancing Translational Sciences (NCATS), and Sage Bionetworks with support from the Neurofibromatosis Therapeutic Acceleration Program (NTAP) at Johns Hopkins, have developed and characterized a set of immortalized patient derived plexiform neurofibroma Schwann cells in an initial effort to create a series of cells that represent the genetic diversity of human plexiform tumors.1 

Through generous support from the Neurofibromatosis Therapeutic Acceleration Program (NTAP), purchasers of these cells are eligible to receive up to a 100% reimbursement from NTAP.  For additional information and to apply for reimbursement after a purchase, please contact NTAP at info@n-tap.org

Through exogenous expression of human telomerase reverse transcriptase (hTERT) and murine cyclin-dependent kinase (mCdk4) using retroviral (and subsequently lentiviral) vectors carrying the hTERT and mCdk4 genes, normal (NF1 wild-type, +/+), neurofibroma-derived Schwann cells that were heterozygous (+/-) for NF1 mutation, and homozygous (-/-) for NF1 mutation, were immortalized. 1

This product, and other immortalized cells from the collection, have undergone in-depth characterization and authentication.

Real time PCR for hTERT and mCdk4 Proliferation rate
NF1 mutational analysis Doubling time
Karyotyping Apoptosis
Cell authentication Microsatellite instability
Contact inhibition Tumorigenicity

 

To learn more and view characterization data, go to: https://www.synapse.org/#!Synapse:syn7239479/wiki/405899

Gosline, SJC Synapse http://doi.org/10.7303/syn4939906 (2017).
Gosline, SJC Synapse http://doi.org/10.7303/syn4940963 (2017).
Gosline, SJC Synapse http://doi.org/10.7303/syn4939874 (2017).
NCBI Sequence Read Archive SRP125359 (2018)
  • Mixture of 6 single-cell clones derived from P16 immortalized primary culture of plexiform neurofibroma cells (ipNF05.5)
  • Tumor: homozygous disease (NF1-/-), plexiform neurofibroma
  • Oncogenes: transduced with mCdk4 cDNA and hTERT
  • Neurofibroma Schwann cell culture transduced with human telomerase (TERT) cDNA and mouse Cdk4 cDNA in lentivirus vectors (no selection), to immortalize the cells.  
Proliferation rate: 82% BrdU positive at 75% confluency
Complete Growth Medium

The basal medium for this cell line is Dulbecco's Modified Eagle's Medium (DMEM; ATCC 30-2002). To make the complete medium add the following components to 500 mL DMEM:

  • 56 mL FBS (ATCC 30-2020) 
  • 5.6 mL L-glutamine from stock 200mM (ATCC 30-2214)
Subculturing
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.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
  3. Add 2.0 to 3.0 mL of Trypsin-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. Add appropriate aliquots of the cell suspension to new culture vessels.
    Cultures can be established between 1.5 x 104 and 3.0 x 104 viable cells/cm2.
  6. Incubate cultures at 37°C.
Interval: Maintain cultures at a cell concentration between 8.0 X 103 and 1.9 X 105 cell/cm2.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:8 is recommended
Medium Renewal: 2 to 3 times per week
Cryopreservation Culture media + 10% DMSO (ATCC 4-X)
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
Cells per Vial Approximately 2.0 to 3.0 x 106 cells
Volume 1.0 mL
STR Profile
Amelogenin: X,Y
CSF1PO: 10,11
D13S317: 11
D16S539: 13,14
D5S818: 11
D7S820: 10,12
THO1: 8,9
TPOX: 8,11
vWA: 16
Sterility Tests Bacteria and yeast: No growth
Mycoplasma: No growth
Viral Testing Hepatitis B: None detected
Cytomegalovirus: None detected
Human immunodeficiency virus: None detected
Epstein-Barr virus: None detected
Human papillomavirus: None detected
Population Doubling Time Per depositor about 33.5 hours
Name of Depositor M Wallace, University of Florida, College of Medicine, Gainesville, FL
Year of Origin 2008
References

Li H, et al. Immortalization of Human Normal and NF1 Neurofibroma Schwann Cells. Lab Invest 96(10):1105-15, 2016. PubMed: 27617404

Ryman DA, et al. Pharmacological and genomic profiling of neurofibromatosis type 1 plexiform neurofibroma-derived schwann cells. Sci Data 5:180106, 2018. PubMed: 29893754

Kraniak JM, et al. Development of 3D culture models of plexiform neurofibroma and initial application for phenotypic characterization and drug screening. Exp Neurol 299(Pt B):289-298, 2018. PubMed: 29055717

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
Other Documentation
Restrictions

For commercial accounts, this cell line is only distributed under the terms of a fully signed and executed ATCC® Material Transfer Agreement and Addendum. If the commercial account is screening per completed Addendum, the recipient will be required to pay a Screening Fee (ATCC® ACS-2103F™).

Screening Use is defined as use of Biological Material in small molecule and biologic drug discovery, including initial target identification and validation, assay development, high throughput screening, hit identification, lead optimization, and selection of candidates for clinical development.

If the commercial account is not screening per the completed Addendum, the recipient will not be required to pay a Screening Fee.

References

Li H, et al. Immortalization of Human Normal and NF1 Neurofibroma Schwann Cells. Lab Invest 96(10):1105-15, 2016. PubMed: 27617404

Ryman DA, et al. Pharmacological and genomic profiling of neurofibromatosis type 1 plexiform neurofibroma-derived schwann cells. Sci Data 5:180106, 2018. PubMed: 29893754

Kraniak JM, et al. Development of 3D culture models of plexiform neurofibroma and initial application for phenotypic characterization and drug screening. Exp Neurol 299(Pt B):289-298, 2018. PubMed: 29055717