Original cell line stocks for HCC38, HCC1143, HCC1187, HCC1954, HCC1937 and HCC1395 were obtained directly from the American Type Culture Collection (ATCC), cultured according to Neve et al [17 (link)], and passaged minimally prior to harvesting for the present study. For each respective cell line, normal matched DNA derived from B lymphoblastoid cell lines (HCC38 BL, HCC1143 BL, HCC1187 BL, HCC1954 BL, HCC1937 BL, HCC1395 BL) were purchased from ATCC. Breast cancer c ells were harvested at approximately 75% confluency and genomic DNA was isolated using DNeasy Blood and Tissue Kit (Qiagen) according to the standard manufacturer protocol. The concentration of DNA was measured with the ND-1000 NanoDrop spectrophotometer (NanoDrop Technologies).
Hcc1954 bl
Manufactured by American Type Culture Collection
The HCC1954 BL is a cell line derived from a human breast cancer sample. It serves as a widely used model for breast cancer research.
Automatically generated - may contain errors
Lab products found in correlation
Hcc1395, by American Type Culture Collection (3 mentions)
Hcc1954, by American Type Culture Collection (3 mentions)
Hcc 1143 bl, by American Type Culture Collection (2 mentions)
Hcc38 bl, by American Type Culture Collection (2 mentions)
Hcc1937, by American Type Culture Collection (2 mentions)
Hcc 1187bl, by American Type Culture Collection (1 mentions)
Nanodrop nd 1000 spectrophotometer, by Thermo Fisher Scientific (1 mentions)
Dneasy blood and tissue kit, by Qiagen (1 mentions)
Hcc38, by American Type Culture Collection (1 mentions)
Hcc1187, by American Type Culture Collection (1 mentions)
Hcc1143, by American Type Culture Collection (1 mentions)
H2009, by American Type Culture Collection (1 mentions)
H1437, by American Type Culture Collection (1 mentions)
Monarch hmw dna extraction kit for tissue, by New England Biolabs (1 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
Automated cell counter, by Bio-Rad (1 mentions)
3 protocols using hcc1954 bl
1
Breast Cancer Cell Line DNA Extraction
2
High-quality DNA Extraction from Cell Lines
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Cell lines were purchased from the American Type Culture Collection (ATCC). All cell lines were cultured according to ATCC handling guidelines at 37°C and 5% CO2. Cell counts were enumerated by Automated Cell Counter (BioRad, TC20), washed twice with Phosphate Buffered Saline (Gibco, 10010023), placed into 6 × 106 cell aliquots, flash frozen in liquid nitrogen, and stored at −80°C. Cancer cell lines: HCC1954 (ATCC, CRL-2338), H2009 (ATCC, CRL-5911), H1437 (ATCC, CRL-5872), and HCC1395 (ATCC, CRL-2324). Normal cell lines: HCC1954 BL (ATCC, CRL-2339), BL2009 (ATCC, CRL-5961), HCC1937, BL1437 (ATCC, CRL-5958), and HCC1395 BL (ATCC, CRL-2325). Utilizing the Monarch® HMW DNA Extraction Kit for Tissue (New England Biolabs, T3060), high molecular weight (HMW) DNA was extracted from 6 × 106 cells for all cell lines.
3
Preparation and Manipulation of B Lymphoblast Cell Lines
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Cell lines HCC1143BL, HCC1954BL, HCC1395BL, HCC38BL, and HCC1954 (Gazdar et al., 1998) were purchased from ATCC and cultured in RPMI-1640 containing penicillin-streptomycin and 10% FBS. For experiments, B lymphoblast cell lines were used at a concentration of approximately 1E6 cells/ml.
For certain experiments using B lymphoblasts, cells were pre-treated with protein degradation inhibitors or vehicle for times indicated, and cell surface MHC Class I removed by acid stripping. This was accomplished by bathing approximately 2E7 cells in 10 mL acid stripping buffer (0.132M citric acid, 0.06M sodium phosphate, pH 3.0) for 2 min on ice, followed by neutralization in 40 mL ice cold medium. Cells were washed once in medium prior to experimental treatment.
Cells were collected for mass spec by centrifugation, which was preceded by trypsin detachment for adherent cell lines (HCC1954). Cell pellets were washed once in PBS and cell count and viability was assessed using a Vi-CELL XR.
For certain experiments using B lymphoblasts, cells were pre-treated with protein degradation inhibitors or vehicle for times indicated, and cell surface MHC Class I removed by acid stripping. This was accomplished by bathing approximately 2E7 cells in 10 mL acid stripping buffer (0.132M citric acid, 0.06M sodium phosphate, pH 3.0) for 2 min on ice, followed by neutralization in 40 mL ice cold medium. Cells were washed once in medium prior to experimental treatment.
Cells were collected for mass spec by centrifugation, which was preceded by trypsin detachment for adherent cell lines (HCC1954). Cell pellets were washed once in PBS and cell count and viability was assessed using a Vi-CELL XR.
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