TempoMito™
Mitochondrial Biosensor Assay
TempoMito™
Mitochondrial Biosensor Assay
TempoMito™ mitochondrial calcium biosensor responds as a fluorescent reporter: 605nm-625nm fluorescent range. It is incorporated into Tempo’s proprietary immortalized human neuroepithelial cell line, TempoRapidTM. TempoMito™ control cells serve as a control for all products containing the TempoMito™ biosensor. This product can be used for single-cell applications, as well as medium to high-throughput screening platforms.
TempoMito™-HEK293t
TempoMito™-HEK293t Cell Line
HEK293t cell line is one of the most widely used human immortalized cell lines in research and drug development. The cells are very easy to work with and can perform a variety of functional applications (e.g., protein purification, expression studies, differential screening, functional reporter assays, receptor-ligand assays, etc).
TempoMito™ biosensor is an mitochondrial calcium flux sensor. It is introduced to the cells using proprietary liposome mediated methods and is expressed in the mitochondria (using marker proteins such as COX IV). HEK293t stably expressing TempoMito™ biosensor has been developed for monitoring mitochondrial calcium flux and cytotoxicity using a highly sensitive fluorescent based biosensor as reporter.
Culture Medium: DMEM with 2mM L-Glutamine; 10% Fetal Bovine Serum (FBS). Subculture Routine: Split sub-confluent using 0.025% trypsin/EDTA solution. Allow the flask to sit at 37°C until the cells detach. Add fresh medium, aspirate and dispense into new culture flasks. Passage ratio: 1-to-9.
TempoMito™-U2OS
TempoMito™-U2OS Cell Line
U2-OS cell line, originally known as the 2T cell line, is an immortalized human cancer cell line. It is widely used in research and drug development, as it offers a variety of application potentials.
TempoMito™ biosensor is an mitochondrial calcium flux sensor. It is introduced to the cells using proprietary liposome mediated methods and is expressed in the mitochondria (using marker proteins such as COX IV). U2-OS stably expressing TempoMito™ biosensor has been developed for monitoring mitochondrial calcium flux and cytotoxicity using a highly sensitive fluorescent based biosensor as reporter.
Culture Medium: McCoy′s 5a medium with 1.5 mM Glutamine or DMEM with 2mM L-Glutamine; 10% Fetal Bovine Serum (FBS). Subculture Routine: Split sub-confluent using 0.025% trypsin/EDTA solution. Allow the flask to sit at 37°C until the cells detach. Add fresh medium, aspirate and dispense into new culture flasks. Passage ratio: 1-to-6
TempoMito™ + Tempo-iAstro™
TempoMito™ Mitochondrial Biosensor Assay using Human Astrocytes
TempoMito™-iAstro cells contain mitochondrial biosensors in human iPS-derived astrocytes. Disruption of mitochondrial functions in human astrocytes has been implicated in Alzheimer’s disorder and ischemia. Tempo’s biosensor responds to mitochondrial calcium changes and reports in the 605nm-625nm fluorescent range (Excitation/Emission) as calcium levels increase or decrease. With excellent signal-to-noise ratios, these sensors can operate on second-to-minute scales. This tool provides an efficient way of detecting changes in the mitochondrial organelle and enables high-throughput screening approaches for drug discovery and toxicology screening.
TempoMito™-iAstro: Mitochondrial Calcium Biosensor Assay using human Astrocytes
TempoMito™-iAstro are derived from integration-free induced pluripotent stem cell (iPSC) lines under a fully defined proprietary astrocytes induction condition. TempoMito™-iAstro cells are plated as monolayers in culture and express markers such as GFAP. TempoMito™ biosensor is introduced to the cells using proprietary liposome mediated methods and is expressed in the mitochondria (using marker proteins such as COX IV).
TempoMito™ + Tempo iNStem™
TempoMito™ Mitochondrial Biosensor Assay using Human Neural Progenitors
TempoMito™-iNStem cells comprise mitochondrial biosensors incorporated into human iPS-derived neural progenitor cells. Mitochondrial dysfunctions have been implicated in human neurological and psychiatric disorders, and neural progenitor cells have been used as a cellular model. Tempo’s biosensor responds to mitochondrial calcium changes and reports in the 605nm-625nm fluorescent range (Excitation/Emission) as calcium levels increase or decrease. With excellent signal-to-noise ratios, these sensors can operate on second-to-minute scales. This tool provides an efficient way of detecting changes in the mitochondrial organelle and enables high-throughput screening approaches for drug discovery and toxicology screening.
TempoMito™-iNStem;: Mitochondrial Calcium Biosensor Assay using human Neural Stem Cells. They are derived from integration-free induced pluripotent stem cell (iPSC) lines under a fully defined proprietary neural induction condition. TempoMito™-iNStem cells are polarized structures when plated as a monolayer in culture and express markers such as Vimentin. We’ve validated iNStem™ cells to be multipotent neural and glial progenitor cells, as they can be further differentiated into neural and glial cell types under specific in vitro reprogramming conditions. TempoMito biosensor is introduced to the cells using proprietary liposome mediated methods and is expressed in the mitochondria (using marker proteins such as COX IV).
TempoMito™ + TempoRapid™
TempoMito™ Mitochondrial Biosensor Assay using Immortalized Human Neuroepithelial Cells
TempoMito™ mitochondrial calcium biosensor responds as a fluorescent reporter: 605nm-625nm fluorescent range. It is incorporated into Tempo’s proprietary immortalized human neuroepithelial cell line, TempoRapidTM. TempoMito™ control cells serve as a control for all products containing the TempoMito™ biosensor. This product can be used for single-cell applications, as well as medium to high-throughput screening platforms.
Applications
TempoATP™-TempoMito™ incorporated cells are intended for research and development, compound discovery and therapeutics development use only. It is not a product for human testing or diagnostics.
Neurotoxicity testing for reporting cellular calcium fluctuations
Phenotypic Assays (i.e., mitochondrial toxicity and function)
Chemical compound or small molecules testing
Cell morphology or cell death research studies
High Content Assays
Live Cell Imaging
Research and development on animal models for human diseases
Studying neural maturation, networking and function, and cations-mediated pathways
Mitochondrial and Cellular metabolism
Receptor studies (e.g., IGF-I, IGF-II)
Specifications
~1×10^6 cells per 1ml of freezing medium (vial)
Long-term Storage: liquid nitrogen
Growth Properties: adherent
Technology used: an in-house developed proprietary feeder-free, serum-free, viral/genetic-elements-free, integration-free reprogramming.
QC: Sterility, Safety (BioSafety Level 2), HIV/viruses, bacteria, fungi: negative. Cell viability post-thawing (>90%)
TempoMito™-HEK293t SKU302
1 vial = ~0.5×10^6 cells
TempoMito™-U2OS SKU301
1 vial = ~0.5×10^6 cells
TempoMito™-iAstro SKU103
TempoMito™-iNStem SKU107
Storage: remove cryovials (dry ice packaging) and place the vial into liquid nitrogen for storage. Alternatively, thaw and use the cells immediately.
TempoMito™-Rapid SKU109
Storage: remove cryovials (dry ice packaging) and place the vial into liquid nitrogen for storage. Alternatively, thaw and use the cells immediately.
Starting Materials: immortalized human cell line of neuro-epithelial origin. Cells should grow fairly well (doubling time <36 h) without passage limitation in a conventional medium DMEM or DMEM/F12 media with a maximum of 10% serum and selection antibiotics (G418, hygromycin or puromycin) at reasonable concentrations.
Frequently Requested Cell Lines
The following are disease cell models that we are able to create through TempoMito™-HEK293t and TempoMito™-U2OS
Cell Line Name | Histology Type | |
U2OS | Bone carcinoma | |
HEK293 | Immortalized Kidney Epithelial | |
SW 480 | Colorectal adenocarcinoma | |
U87MG | Brain cancer | |
Hep G2 | Hepatocellular carcinoma | |
NIH3T3 | Embroyonic fibroblast, murine | |
HT1080 | Carcinoma (connective tissue) | |
HCT116 | Colorectal cancer | |
MCF 7 | Breast cancer | |
SW 1990 | Pancreatic cancer | |
NCI-H2126 | Non-Small cell cancer | |
HUVEC | Umbilical vascular endothelium | |
HeLa | Cervical epithelial adenocarcinoma | |
CG4 | Glial cell line (from rat) | |
A-204 | Muscle epithelial | |
A549 | Lung epithelial cell line | |
BT-142 | Oligoastrocytoma, suspension | |
NK-92 | Peripheral blood, natural killer cells | |
CHO | Chinese hamster ovarian cell line | |
NCI-H69 | Lung tumor cell aggregate, suspension | |
ARPE-19 | Retinal pigmented epithelial | |
Beta-TC-6 | Beta cell (mouse) | |
BxPC-3 | Pancreatic epithelial, adenocarcinoma | |
Jurkat cells | Peripheral T lymphocyte, suspension | |
SW 780 | Urinary bladder epithelial carcinoma | |
MeT-5A | Mesothelium, from non-cancerous | |
NFKB reporters | Email us | |
Mesenchymal stromal cells | Email us | |
Chemokine Reporter cells | Email us | |
cell line of your choice | Email us |
References
TempoMito™-HEK293t
- Dykens JA, Jamieson JD, Marroquin LD, Nadanaciva S, Xu JJ, Dunn MC, Smith AR, Will Y. Toxicol Sci. 2008 Jun;103(2):335-45.
- Dykens JA, Will Y. Drug Discov today. 2007 Sep; 12 (17-18): 777-85.
TempoMito™-iAstro
- Martin LJ. Prog Mol Biol Transl Sci. 2012;107:355-415.
- Martin LJ. Pharmaceuticals (Basel). 2010;3(4):839-915.
- Chen K, Northington FJ, Martin LJ. Brain Struct Funct. 2010 Mar;214(2-3):219-34.
- Palomo GM, Manfredi G. Brain Res. 2015 May 14;1607:36-46.
- Dykens JA, Jamieson JD, Marroquin LD, Nadanaciva S, Xu JJ, Dunn MC, Smith AR, Will Y. Toxicol Sci. 2008 Jun;103(2):335-45.
- Dykens JA, Will Y. Drug Discov today. 2007 Sep; 12 (17-18): 777-85.
TempoMito™-iNStem
- Martin LJ. Prog Mol Biol Transl Sci. 2012;107:355-415.
- Martin LJ. Pharmaceuticals (Basel). 2010;3(4):839-915.
- Chen K, Northington FJ, Martin LJ. Brain Struct Funct. 2010 Mar;214(2-3):219-34.
- Palomo GM, Manfredi G. Brain Res. 2015 May 14;1607:36-46.