HT-1080 LifeAct-TagGFP2 Stable Cell Line
A stable LifeAct-TagGFP2 expressing human fibrosarcoma cell line
- Long-term actin visualization - 25 passages guaranteed
- Characteristics proven to be identical to the parental HT-1080 cell line
- LifeAct technology guarantees no interference with cytoskeletal dynamics
This cell line was developed in cooperation with
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|40101||HT-1080 LifeAct-TagGFP2: HT-1080 cells expressing LifeAct-TagGFP2, 5x105 cells/vial||1|
To create the stable LifeAct-TagGFP2 expressing cell line, human fibrosarcoma cells were stably transfected with a plasmid coding for TagGFP2-labeled LifeAct .
The HT-1080 LifeAct-TagGFP2 cells present highly dynamic filamentous actin. As a result of stable LifeAct expression, perfect visualization of filamentous actin with very low background fluorescence is achieved without any interference to cellular dynamics. Moreover, full characterization of the cells revealed that they displayed identical behavior to wild type (wt) cells.
Due to the cell line’s fast growth and the absence of contact inhibition, it constitutes an ideal tumor model. These characteristics combined with bright and specific F-actin staining lead to a powerful tool with versatile applications in live cell imaging and high-content approaches.
*For further information on TagGFP2, please refer to www.evrogen.com.
|Organism||Homo sapiens (human)|
|Source||Connective tissue disease: fibrosarcoma|
Basic Characterization of HT-1080 LifeAct-TagGFP2
Genomic authentication of human cell lines by means of short tandem repeat (STR) analysis is widely used to assure cell line identity and to investigate cell line purity.
Short tandem repeat polymorphisms in the human genome are used to differentiate between individuals. By identifying the short repeats of a specific sequence at typical locations in the genome, it is possible to create a genetic profile of an individual human being (as used in forensics), or of a cell line.
ibidi’s HT-1080 LifeAct-TagGFP2 cells were analyzed using the above mentioned method. The analysis was performed by DSMZ, an ISO-certified Leibniz institute. The analysis revealed that HT-1080 LifeAct-TagGFP2 cells are identical to HT-1080 wild type cells.
Cell Proliferation Analysis:
HT-1080 LifeAct-TagGFP2 and wild type cells were cultured for 8 days using the same defined conditions as the cultures’ start points, and then analyzed for cell number each day.
Results from this experiment showed that both cell lines grew at equal rates over the predefined period of time. This result was confirmed using statistical tests.
Fluorescence Intensity in Cell Population:
HT-1080 LifeAct-TagGFP2 and wild type cells were analyzed for fluorescence intensity and number of positive cells using flow cytometry.
At least 95 % of the cells were fluorescent with on average a two log shift in fluorescence intensity compared to wild type cells.
Functional Characterization of HT-1080 LifeAct-TagGFP2
Chemotaxis assays of HT-1080 LifeAct-TagGFP2 and wild type cells were analyzed in µ-Slides Chemotaxis 2D and µ-Slides Chemotaxis 3D, using fetal calf serum (FCS) as a chemoattractant (see Application Notes 14 and 17 for further details).
Results from these assays showed that HT-1080 LifeAct-TagGFP2 cells migrate with the same speed and forward migration index (FMI) as wild type cells. These results were confirmed using the Student’s t-test. p < 0.05 was considered significant.
HT-1080 LifeAct-TagGFP2 and wild type cells were seeded in a µ-Slide 8 well combined with a Culture-Insert. After 24 hours of video microscopy, wound closure was analyzed using the WimScratch automated software solution.
HT-1080 LifeAct-TagGFP2 and wild type cells showed comparable cell front velocities. These results were confirmed using the Student’s t-test. p < 0.05 was considered significant.
More stable LifeAct transfected cell lines will be available in the future. Please contact ibidi for detailed information.
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