產(chǎn)品名稱(chēng) |
BT142 mut/- |
商品貨號(hào) |
B228265 |
Organism |
Homo sapiens, human |
Tissue |
Brain |
Cell Type |
Neural |
Product Format |
frozen |
Morphology |
Neurosphere |
Culture Properties |
Suspension |
Biosafety Level |
1 [It is the responsibility of the investigator to determine appropriate safety procedures for use with this material. As a reference, laboratory safety is discussed in the publication Biosafety in Microbiological and Biomedical Laboratories and can be accessed by searching "BMBL" at www.cdc.gov.]
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 |
Oligoastrocytoma Grade III |
Age |
38 years |
Gender |
Male |
Ethnicity |
Caucasian |
Storage Conditions |
Liquid Nitrogen Vapor Phase |
Images |
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Clinical Data |
38 years
Male
Caucasian
Oligoastrocytoma Grade III |
Comments |
Point mutations in isocitrate dehydrogenase I (IDH1) and IDH2 are found in majority of grade II and III gliomas. R132H is the most common IDH1 substitution found in gliomas.
BT142 mut/- contains a homozygous IDH1 R132H mutation, which originated from a heterozygous IDH1 R132H BT142 cells.
The cells grow as phase-bright, smooth spheres.
The neurospheres should not get too big, ragged or dark as this is a sign of unhealthy, dying cells.
The cells should be passaged when the neurospheres are 200 to 400 µm in size. |
Complete Growth Medium |
There are two options for the base medium for this cell line.
Option 1: NeuroCult NS-A Proliferation kit (Catalog No. 5751, Stem Cell Technologies)
Option 2: DMEM/F12 (1:1) (Catalog No. 30-2006, ATCC) with an additional 0.9% glucose, 4 mM L-glutamine (Catalog No. 30-2214, ATCC), 25 µg/mL insulin, 100 µg/mL transferrin, 20 nM progesterone, 15 µM putrescine and 30 nM selenite
To make the complete growth medium, add the following supplements to either options of the base medium (see above):
- 20ng/mL recombinant human Epidermal Growth Factor (EGF, Catalog No. 100-15, PeproTech)
- 100 ng/mL recombinant human Platelet-Derived Growth Factor-AA (PDGF-AA, Catalog No. 100-13A, PeproTech)
- 20 ng/mL recombinant human Fibroblast Growth Factor (R&D Systems, Catalog No. 233-FB)
- 2 µg/mL heparan sulfate (Catalog No. H3149, Sigma)
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Subculturing |
The cells grow as phase-bright, smooth spheres. The neurospheres should not get too big, ragged or dark as this is a sign of unhealthy, dying cells. The cells should be passaged when the neurospheres are about 200-400 μm in size.
Volumes used in this protocol are for 75 cm2 flask.
- Harvest and collect the entire cell suspension from the culture flask into a 15 mL tube.
- Centrifuge at 200 x g for 10 minutes.
- Aspirate supernatant, leaving approximately 200 µL to cover the pellet.
- Add 1 mL of complete culture medium.
- Triturate cells with a P1000 micropipette set to 800 µL by pipetting up and down 40 times or until the cells appear to be in a single cell suspension.
- Add 8 mL of complete culture medium and centrifuge at 200 x g for 10 minutes.
- Aspirate supernatant and resuspend the cells in 2 mL of complete culture medium.
- Count viable cells using trypan blue exclusion assay on a hemacytometer.
- Seed single cells between ranges of 8 x 103 to 2 x 104 cells/cm2.
Note: If accurate cell count is necessary, Accumax (Catalog No. AM105, Innovative Cell Technologies) can be used; however, the cells may take some time to recover from an enzymatic dissociation.
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Cryopreservation |
Freeze medium: Complete growth medium (90%) supplemented with 10% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase |
Name of Depositor |
This cell line was deposited by Samuel Weiss, Ph.D. and Gregory Cairncross, Ph.D. of the University of Calgary |
References |
Luchman HA et al. An in vivo patient-derived model of endogenous IDH1-mutant glioma. Neuro Oncol. 14:184-191, 2012. PubMed: 22166263
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