Cell Culturing: Difference between revisions
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=Cell Culturing= | =Cell Culturing= | ||
=HeLa= | ==HeLa== | ||
[[Image: | [[Image:HeLa.jpg|200px|right]] | ||
Derived from Henrietta Lacks in 1951, HeLa is an immortal cell line that is useful for its remarkable prolificity and durability. It was the inspiration for the national bestseller, The Immortal Life of Henrietta Lacks, written by the incredibly attractive Rebecca Skloots. | Derived from Henrietta Lacks in 1951, HeLa is an immortal cell line that is useful for its remarkable prolificity and durability. It was the inspiration for the national bestseller, The Immortal Life of Henrietta Lacks, written by the incredibly attractive Rebecca Skloots. | ||
Because of their high reproductive rate, HeLa cells are a good beginner transfection cell line to test the efficacy of the CPNP delivery of the origami. | |||
*Culture HeLa in 75 cm<sup>2</sup> flasks in a humidified incubator at 37°C and 5% CO<sub>2</sub> in DMEM supplemented with 10% FBS and 1% Streptomycin/Penicillin. Cells were mantained at subconfluency and passaged every three days. | |||
*Prior to the day of transfection, cells were seeded on 12-well plate at 1 x 10<sup>5</sup> cells/well and were allowed to incubate for 24 hours. | |||
*On day of transfection, the CPNP/origami nanoparticles were added to each well as described in [[OhioMod2013:Results|results]]. | |||
==HEK-293== | |||
HEK-293 cells are kidney cells obtained in 1970 that were [http://en.wikipedia.org/wiki/Transformation_%28genetics%29 transformed] with an adenovirus to form a cell line that is easily cultured and transfected. HEK-293 is commonly used in calcium phosphate transfections as well as (with the 293T variant) propagation of adenoviral vectors. | |||
We expect to use HEK-293 as a positive control, especially if we use the SV40 promoter sequence on the origami. | |||
Latest revision as of 09:30, 7 May 2013
Cell Culturing
HeLa
Derived from Henrietta Lacks in 1951, HeLa is an immortal cell line that is useful for its remarkable prolificity and durability. It was the inspiration for the national bestseller, The Immortal Life of Henrietta Lacks, written by the incredibly attractive Rebecca Skloots.
Because of their high reproductive rate, HeLa cells are a good beginner transfection cell line to test the efficacy of the CPNP delivery of the origami.
- Culture HeLa in 75 cm2 flasks in a humidified incubator at 37°C and 5% CO2 in DMEM supplemented with 10% FBS and 1% Streptomycin/Penicillin. Cells were mantained at subconfluency and passaged every three days.
- Prior to the day of transfection, cells were seeded on 12-well plate at 1 x 105 cells/well and were allowed to incubate for 24 hours.
- On day of transfection, the CPNP/origami nanoparticles were added to each well as described in results.
HEK-293
HEK-293 cells are kidney cells obtained in 1970 that were transformed with an adenovirus to form a cell line that is easily cultured and transfected. HEK-293 is commonly used in calcium phosphate transfections as well as (with the 293T variant) propagation of adenoviral vectors.
We expect to use HEK-293 as a positive control, especially if we use the SV40 promoter sequence on the origami.
