BT89 (ATCC® CRL-3411)

Organism: Homo sapiens, human  /  Tissue: brain  /  Disease: glioblastoma

Organism Homo sapiens, human
Tissue brain
Product Format frozen 1.0 mL
Morphology bright smooth spheres
Culture Properties Aggregate/Suspension
Note: Neurospheres must be mechanically broken up when they get too large (they form ragged edges and dark areas within the neurospheres). Do not dispense cells into single cell suspension during this process, but just reduce in size of clusters to maintain a healthy state.
Biosafety Level 1

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 glioblastoma
Age 44
Gender male
Applications

A model of glioblastoma; useful for drug discovery and disease modeling.

Images
Comments MGMT promoter methylation Methylated, EGFR wild-type, PTEN heterozygous mutant, TP53 wild-type, IDH wild-type
Complete Growth Medium NeuroCult Basal Medium w/ proliferation Supplement + 20 ng/mL EGF + 20 ng/mL β-FGF + 2 μg/mL Heparin sulphate. Protect from light and use complete medium within 10 days.

NeuroCult NS-A Proliferation Kit (Stem Cell Technologies cat# 5751)

  • 450 mL NeuroCult Basal Medium (SCT cat# 5750)
  • 50 mL NeuroCult Supplement (SCT cat# 5753)

20 μg/mL EGF: use 1 μL/1 mL culture medium. To prepare 20 μg/mL EGF stock solution, aseptically combine:

  • 100 μg EGF (PeproTech cat# 100-15)
  • 1 mL PBS (ATCC 30-2200)

Then further dilute by adding 4 mL PBS/0.1% BSA

20 μg/mL β-FGF: use 1 μL/1 mL culture medium. To prepare 20 μg/mL EGF stock solution, aseptically combine:

  • β-FGF (PeproTech cat# 100-18B)
  • 1 mL 5 mM Tris PH 7.6

Then further dilute by adding 4 mL PBS/0.1% BSA

Store in working aliquots at -20°C. EGF and β-FGF is stable for 1 year when prepared and stored as directed. Do not repeat freeze thaw.

2 mg/mL Heparin Solution (Stem Cell Technologies cat# 07980): use 1 μL/1 mL culture medium. Store solution at 4°C.

Subculturing
Cultures can be maintained by addition or replacement of fresh medium. Start cultures at 2.0 X 105 cells/mL and maintain between 1.0 X 105 and 3.0 X 106 cells/mL.

Medium Renewal: Add fresh medium every 2 to 3 days (depending on cell density).
Cryopreservation Complete culture medium(with or without EGF/╬▓-FGF/Heparin) + 10% DMSO
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
Cells per Vial Approximately 2.0 to 3.0 x 106 viable cells
Volume 1.0 mL
STR Profile
Amelogenin: X,Y
CSF1PO: 12,13
D13S317: 9,11
D16S539: 12
D5S818: 12
D7S820: 10,12
THO1: 7
TPOX: 8
vWA: 16,18
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
Name of Depositor University of Calgary
Year of Origin 2007
References

Cusulin C, et al. Precursor States of Brain Tumor Initiating Cell Lines Are Predictive of Survival in Xenografts and Associated with Glioblastoma Subtypes. Stem Cell Reports 5(1): 1-9, 2015. PubMed: 26095605

Kelly JJ, et al. Proliferation of human glioblastoma stem cells occurs independently of exogenous mitogens. Stem Cells 27(8): 1722-33, 2009. PubMed: 19544433

Luchman A, et al. Dual mTORC1/2 Blockade Inhibits Glioblastoma Brain Tumor Intiating Cells In Vitro and In Vivo and Synergizes with Temozolomide to Increase Orthotopic Xenograft Survival. Clin Cancer Res 20(22): 5756-67, 2014. PubMed: 25316808

Stechishin O, et al. On-target JAK2/STAT3 inhibition slows disease progression in orthotopic xenografts of human glioblastoma brain tumor stem cells. Neuro Oncol 15(2): 198-207, 2013. PubMed: 23262510

Basic Documentation
Other Documentation
References

Cusulin C, et al. Precursor States of Brain Tumor Initiating Cell Lines Are Predictive of Survival in Xenografts and Associated with Glioblastoma Subtypes. Stem Cell Reports 5(1): 1-9, 2015. PubMed: 26095605

Kelly JJ, et al. Proliferation of human glioblastoma stem cells occurs independently of exogenous mitogens. Stem Cells 27(8): 1722-33, 2009. PubMed: 19544433

Luchman A, et al. Dual mTORC1/2 Blockade Inhibits Glioblastoma Brain Tumor Intiating Cells In Vitro and In Vivo and Synergizes with Temozolomide to Increase Orthotopic Xenograft Survival. Clin Cancer Res 20(22): 5756-67, 2014. PubMed: 25316808

Stechishin O, et al. On-target JAK2/STAT3 inhibition slows disease progression in orthotopic xenografts of human glioblastoma brain tumor stem cells. Neuro Oncol 15(2): 198-207, 2013. PubMed: 23262510