Three different breast cancer cell lines were included in this study to represent a variety of phenotypes: MDA-MB-231 (triple-negative, but expected to express EGFR; ATCC® HTB-26™), MDA-MB-468 (triple-negative, but over-expresses EGFR; ATCC® HTB-132™), and AU565 (over-expresses HER2, and expected to express EGFR; ATCC® CRL-2351™). All cell lines were incubated for the duration of experiments in 37 °C and 5% CO2 level. Dulbecco’s modified Eagle’s medium low glucose (DMEM), containing L-glutamine, sodium pyruvate, and glucose was used for MDA-MB-231 and MDA-MB-468 cells. RPMI 1640 medium was used for AU565. All media were supplemented with 10% (v/v) fetal bovine serum, 100 U/mL penicillin and 100 μg/mL streptomycin. Cells were sub-cultured after reaching > 80% confluency and were discarded after ~40 passages.
Crl 2351
Manufactured by American Type Culture Collection
Sourced in Germany, United States
CRL-2351 is a cell line derived from human breast carcinoma tissue. It is a widely used model for breast cancer research.
Automatically generated - may contain errors
Lab products found in correlation
Htb 132, by American Type Culture Collection (2 mentions)
Htb 26, by American Type Culture Collection (2 mentions)
Slide flask, by Thermo Fisher Scientific (1 mentions)
Fetal calf serum (fcs), by Harvard Bioscience (1 mentions)
Ham s f12 media, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Harvard Bioscience (1 mentions)
Trypsin solution, by Corning (1 mentions)
Hcc1806, by American Type Culture Collection (1 mentions)
Hcc1428, by American Type Culture Collection (1 mentions)
Rpmi 1640 medium, by Corning (1 mentions)
Dmem f12 medium, by Merck Group (1 mentions)
Crm htb 26, by American Type Culture Collection (1 mentions)
Htb 22, by American Type Culture Collection (1 mentions)
4 protocols using crl 2351
1
Comparative Analysis of Breast Cancer Cell Lines
2
HER2+ Breast Cancer Cell Line Microgravity Study
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Human breast cancer cells (adenocarcinoma, CRL2351)
were obtained from ATCC© (Wesel, Germany).
All experiments were performed on this commercially
available cell line, so no Ethical Committee approval was
necessary. The cell line is negative estrogen receptor and it
overexpresses the HER2/neu oncogene. The cells were firstly
expanded under 2D-conditions in T125 flasks (Sarstedt,
USA). Ham´s F12-media (Gibco, Germany) supplemented
with 5% fetal calf serum (FCS, Biochrom AG, Germany)
and 1% penicillin/streptomycin (Biochrom, Germany) was
used. The medium was changed three times per week. For
this experiment, 1×106 cells were counted by hemocytometer
and added to six T125 flasks, as the experimental group in the
RPM, and to the same number of flasks for the control group
under 1g conditions. For cytoskeleton staining, the cells were
seeded with a density of 1×105 per cm2 to slide flasks (Thermo
Scientific, Germany).
were obtained from ATCC© (Wesel, Germany).
All experiments were performed on this commercially
available cell line, so no Ethical Committee approval was
necessary. The cell line is negative estrogen receptor and it
overexpresses the HER2/neu oncogene. The cells were firstly
expanded under 2D-conditions in T125 flasks (Sarstedt,
USA). Ham´s F12-media (Gibco, Germany) supplemented
with 5% fetal calf serum (FCS, Biochrom AG, Germany)
and 1% penicillin/streptomycin (Biochrom, Germany) was
used. The medium was changed three times per week. For
this experiment, 1×106 cells were counted by hemocytometer
and added to six T125 flasks, as the experimental group in the
RPM, and to the same number of flasks for the control group
under 1g conditions. For cytoskeleton staining, the cells were
seeded with a density of 1×105 per cm2 to slide flasks (Thermo
Scientific, Germany).
3
Culturing Breast Cancer Cell Lines
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The following breast cancer cell lines, namely triple negative breast cancer (TNBC) HCC1806 (CRL-2335™), ER-positive HCC1428 (CRL-2327™), and HER2-positive AU565 (CRL-2351™) (ATCC, Manassas, VA, USA), were grown in RPMI-1640 medium (Corning, Tewksbury, MA, USA) supplemented with 10% FBS and antibiotics (100 U/mL penicillin, 0.1 mg/mL streptomycin, and 0.25 g/mL amphotericin). MCF10F non-tumorigenic epithelial cells from mammary gland (CRL-10318™, ATCC, Manassas, VA, USA) were cultivated in DMEM/F12 medium (Merck KGaA, Darmstadt, Germany) containing 5% horse serum, 10 ng/mL epidermal growth factor EGF, 10 µg/mL insulin, 0.5 µg/mL hydrocortisone, 100 ng/mL cholera toxin, and mix of antibiotics. Cells were cultured at 37 °C in a controlled humidified atmosphere containing 5% CO2 and passaged with trypsin solution (0.05% trypsin for MCF10F cells and 0.25% trypsin for breast cancer cells, respectively, Corning, Tewksbury, MA, USA).
4
Characterization of Breast Cancer Cell Lines
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A panel of human breast cancer cell lines were selected for this study to represent a variety of characteristics and receptor expression profiles: (i) MDA-MB-231 (MDA231; triple negative, but expected to express EGFR; ATCC ® HTB-26™); (ii) MDA-MB-231, KRas-CRM (MDA231K; same as MDA-MB-231, certified for KRas mutation; ATCC ® CRM-HTB-26™);
(iii) MDA-MB-468 (MDA468; triple negative, but over-expresses EGFR; ATCC ® HTB-132™);
(iv) AU565 (over-expresses HER2, and expected to express EGFR; ATCC ® CRL-2351™); (v) MCF7 (estrogen and progesterone-positive, and expected to express HER2 and EGFR; ATCC ® HTB-22™), and; (vi) MDA-MB-435 (MDA-435): Long known as an estrogen-independent breast cancer cell line expressing HER-2 [25, 26] . MDA-435 cells have been source of controversy since it bears both epithelial and melanocytic markers [27, 28] . All cell lines except AU565 were cultured in DMEM low glucose medium, while AU565 was cultured in RPMI-1640 (both mediums supplemented with 10% (v/v) FBS, 100 U/mL penicillin and 100 μg/ml streptomycin). All cell lines were maintained at 37C and 5% CO 2 and were sub-cultured after reaching 80-90% confluency. Cells were discarded after 40 passages, and frozen cells (stored in liquid nitrogen) were thawed as replacement.
(iii) MDA-MB-468 (MDA468; triple negative, but over-expresses EGFR; ATCC ® HTB-132™);
(iv) AU565 (over-expresses HER2, and expected to express EGFR; ATCC ® CRL-2351™); (v) MCF7 (estrogen and progesterone-positive, and expected to express HER2 and EGFR; ATCC ® HTB-22™), and; (vi) MDA-MB-435 (MDA-435): Long known as an estrogen-independent breast cancer cell line expressing HER-2 [25, 26] . MDA-435 cells have been source of controversy since it bears both epithelial and melanocytic markers [27, 28] . All cell lines except AU565 were cultured in DMEM low glucose medium, while AU565 was cultured in RPMI-1640 (both mediums supplemented with 10% (v/v) FBS, 100 U/mL penicillin and 100 μg/ml streptomycin). All cell lines were maintained at 37C and 5% CO 2 and were sub-cultured after reaching 80-90% confluency. Cells were discarded after 40 passages, and frozen cells (stored in liquid nitrogen) were thawed as replacement.
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