Cultivation of Large Cultures of HeLa Cells in Horse Serum

Perry, Vernon P.; Evans, Virginia J.; Earle, Wilton R.

doi:10.1126/science.121.3153.805

Cited by 14 publications

(9 citation statements)

References 2 publications

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“…HeLa (human cervical squamous carcinoma cells) [21], and CT26 (murine colon adenocarcinoma) [22] were obtained from ATCC (Manassas, VA) and cultured in RPMI1600 medium (Gibco Invitrogen, Carlsbad, CA) with L-glutamine and NaHCO 3 supplemented with heat inactivated fetal bovine serum 10% (vol/vol), penicillin (100 U/mL) and streptomycin (100 µg/mL) (all from Sigma, St Louis, MO) at 37 °C in 5% CO 2 humidified atmosphere in 75 cm 2 flasks (BD Falcon, San Jose, CA). When the cells reached 80% confluence, they were washed with PBS (Sigma) and harvested with 2 mL of 0.25% Trypsin-EDTA solution (Sigma).…”

Section: Methodsmentioning

confidence: 99%

Imidazole metalloporphyrins as photosensitizers for photodynamic therapy: Role of molecular charge, central metal and hydroxyl radical production

et al. 2009

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SUMMARYThe in vitro photodynamic therapy activity of four imidazole-substituted metalloporphyrins has been studied using human (HeLa) and mouse (CT26) cancer cell lines: an anionic Zn porphyrin and a homologous series of three cationic Zn, Pd or InCl porphyrins. A dramatic difference in phototoxicity was found: Pd cationic > InCl cationic > Zn cationic > Zn anionic. HeLa cells were more susceptible than CT26 cells. Induction of apoptosis was demonstrated using a fluorescent caspase assay. The anionic Zn porphyrin localized in lysosomes while the cationic Zn porphyrin localized in lysosomes and mitochondria, as assessed by fluorescence microscopy. Studies using fluorescent probes suggested that the cationic Pd porphyrin produced more hydroxyl radicals as the reactive oxygen species. Thus, the cationic Pd porphyrin has high potential as a photosensitizer and gives insights into characteristics for improved molecular designs.

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Section: Methodsmentioning

confidence: 99%

Imidazole metalloporphyrins as photosensitizers for photodynamic therapy: Role of molecular charge, central metal and hydroxyl radical production

et al. 2009

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“…Cell culture: A human cervical cancer cell line (HeLa) [32] was obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA). The cells were cultured in complete media containing Dulbelcco's modified-eagle medium (DMEM) with l-glutamine and NaHCO 3 , supplemented with 10 % heat-inactivated fetal bovine serum (FBS), and penicillin (100 U mL À1 ) at 378C in a humidified 5 % CO 2 atmosphere in 75 cm 2 flasks (BD Falcon, Franklin Lakes, NJ, USA).…”

Section: Biological Evaluationmentioning

confidence: 99%

Stable Synthetic Bacteriochlorins for Photodynamic Therapy: Role of Dicyano Peripheral Groups, Central Metal Substitution (2H, Zn, Pd), and Cremophor EL Delivery

et al. 2012

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A series of four stable synthetic bacteriochlorins was tested in vitro in HeLa cells for their potential in photodynamic therapy (PDT). The parent bacteriochlorin (BC), dicyano derivative (NC)2BC and corresponding zinc chelate (NC)2BC–Zn and palladium chelate (NC)2BC–Pd were studied. Direct dilution of a solution of bacteriochlorin in an organic solvent (N,N-dimethylacetamide) into serum-containing medium was compared with the dilution of bacteriochlorin in Cremophor EL (CrEL; polyoxyethylene glycerol triricinoleate) micelles into the same medium. CrEL generally reduced aggregation (as indicated by absorption and fluorescence) and increased activity up to tenfold (depending on bacteriochlorin), although it decreased cellular uptake. The order of PDT activity against HeLa human cancer cells after 24 h incubation and illumination with 10 J cm−2 of near-infrared (NIR) light is (NC)2BC–Pd (LD50 = 25 nm) > (NC)2BC > (NC)2BC–Zn ≈ BC. Subcellular localization was determined to be in the endoplasmic reticulum, mitochondria and lysosomes, depending on the bacteriochlorin. (NC)2BC–Pd showed PDT-mediated damage to mitochondria and lysosomes, and the greatest production of hydroxyl radicals as determined using a hydroxyphenylfluorescein probe. The incorporation of cyano substituents provides an excellent motif for the enhancement of the photoactivity and photostability of bacteriochlorins as PDT photosensitizers.

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“…Embryos were disrupted by pipetting and then added to at least a 3_ volume of Dulbecco's modified Eagle's medium containing 10% fetal bovine serum and 1% penicillin/streptomycin. Cells were plated into 75cm 2 flasks. For all the experiments, cells were grown to at most 80% confluence before seeding to 96 well plates and only the cells between passage 3 and passage 8 were employed.…”

Section: Primary Mouse Embryonic Fibroblastsmentioning

confidence: 99%

“…HL-1 cells were a cardiac muscle cell line, first established and characterized the cell line by Dr W. C. Claycomb [2]. Briefly, cells were grown as a monolayer (37°C, 5% CO 2 ) in culture flasks.…”

Section: Mouse Cardiomyocyte Cell Linementioning

confidence: 99%

Comparison of cellular responses induced by low level light in different cell types

et al. 2010

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Discoveries are rapidly being made in multiple laboratories that shed "light" on the fundamental molecular and cellular mechanisms underlying the use of low level light therapy (LLLT) in vitro, in animal models and in clinical practice. Increases in cellular levels of respiration, in cytochrome c oxidase activity, in ATP levels and in cyclic AMP have been found. Increased expression of reactive oxygen species and release of nitric oxide have also been shown. In order for these molecular changes to have a major effect on cell behavior, it is likely that various transcription factors will be activated, possibly via different signal transduction pathways. In this report we compare and contrast the effects of LLLT in vitro on murine embryonic fibroblasts, primary cortical neurons, cardiomyocytes and bone-marrow derived dendritic cells. We also examined two human cell lines, HeLa cancer cells and HaCaT keratinocytes. The effects of 810-nm near-infra-red light delivered at low and high fluences were addressed. Reactive oxygen species generation, transcription factor activation and ATP increases are reported. The data has led to the hypothesis that cells with a high level of mitochondrial activity (mitochondrial membrane potential) have a higher response to light than cells with low mitochondrial activity.

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Cultivation of Large Cultures of HeLa Cells in Horse Serum

Cited by 14 publications

References 2 publications

Imidazole metalloporphyrins as photosensitizers for photodynamic therapy: Role of molecular charge, central metal and hydroxyl radical production

Imidazole metalloporphyrins as photosensitizers for photodynamic therapy: Role of molecular charge, central metal and hydroxyl radical production

Stable Synthetic Bacteriochlorins for Photodynamic Therapy: Role of Dicyano Peripheral Groups, Central Metal Substitution (2H, Zn, Pd), and Cremophor EL Delivery

Comparison of cellular responses induced by low level light in different cell types

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