The KPC-derived PDAC cell line (TB32048) previously generated from a female C57BL/6 KPC mouse was provided as a generous gift from David Tuveson laboratory. TB32048 was cultured for 3–4 passages at maximum 80% confluency in 10% fetal calf-serum (#A15-104, GE Healthcare) in DMEM (#E15-810, PAA) + 100 μg/ml penicillin/streptomycin (#P11-G10, PAA) in a T175 flask in standard conditions (37°C, 5% CO2) and tested regularly in house for mycoplasma (#rep-pt1, Invivogen and #LT07-710, Lonza). Cells were dissociated using 0.1% trypsin (PBS) (#594-18C, Sigma) for 10 min at 37°C and resuspended in PBS and BD Matrigel™ Basement Membrane Matrix High Concentration (#354248, BD Biosciences) in a ratio of 1:1. 1,000 cells in 5 μl was injected into the pancreas using a Hamilton® syringe, 700 series (#10100332, Fisher Scientific). For the experiments used to assess CD138hi cells and anti-CD20 at day 10, 10,000 cells in 30 μl Matrigel, or Matrigel only sham surgery was performed. The peritoneal wall was sutured using 6/0-gauge coated vicryl sutures™ (#W9500T, Ethicon) and skin closed using two 9 mm Clay Adams Clips (#IN015A, VetTech Solutions) and an Autoclip® applier (#IN015B, VetTech Solutions).
Lt07 710
Manufactured by Lonza
The LT07-710 is a laboratory equipment product designed for general laboratory applications. It serves as a versatile tool to support various experimental and analytical procedures. The core function of the LT07-710 is to provide a reliable and consistent platform for laboratory work, without further interpretation or extrapolation on its intended use.
Automatically generated - may contain errors
Lab products found in correlation
Sh3007003, by GE Healthcare (3 mentions)
Sv30010, by GE Healthcare (3 mentions)
Azd6244, by Selleck Chemicals (2 mentions)
Sw620, by American Type Culture Collection (2 mentions)
Rep pt1, by InvivoGen (1 mentions)
Matrigel basement membrane matrix high concentration, by BD (1 mentions)
Rpmi 1640, by Merck Group (1 mentions)
Jurkat, by American Type Culture Collection (1 mentions)
Rs4 11, by American Type Culture Collection (1 mentions)
Nalm6, by American Type Culture Collection (1 mentions)
Molt 4, by American Type Culture Collection (1 mentions)
Advanced dmem, by Thermo Fisher Scientific (1 mentions)
Mia paca 2, by Thermo Fisher Scientific (1 mentions)
S12450, by Thermo Fisher Scientific (1 mentions)
Mia paca 2, by American Type Culture Collection (1 mentions)
Hcc1954, by Thermo Fisher Scientific (1 mentions)
Hct116, by American Type Culture Collection (1 mentions)
Hcc1954, by American Type Culture Collection (1 mentions)
Penicillin and streptomycin, by Thermo Fisher Scientific (1 mentions)
7 protocols using lt07 710
1
Orthotopic Implantation of KPC-derived PDAC Cells
2
Comprehensive Characterization of Cell Lines
3
Comprehensive Characterization of Cell Lines
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Human cancer cell lines, SW620, H358, and H293T, were purchased from ATCC, UT89 was generated from a primary recurrent uterine serous tumor in the laboratory of Dr. Analisa DiFeo. SW620 and H358 were cultured in RPMI, H293T and UT89 were cultured in DMEM. All media was supplemented with 10% FBS (ThermoFisher, SH3007003) and 1% penicillin/streptomycin (GE Healthcare, SV30010). HMECs were obtained from Dr. Mark Jackson. Specimen 48R and derivatives were grown as described previously21 ,22 (link). HMEC derivatives were generated as previously described21 ,23 (link). All cells were grown in a humidified atmosphere containing 5% CO2. All cells lines underwent monthly testing for mycoplasma contamination (Lonza, LT07-710). AZD-6244 (Selleck Chemical, S1008) was dissolved in DMSO at a concentration of 80 mM and stored at −20°C (for in vitro studies). Okadaic Acid (OA) (P212121, LC-O-2220) was dissolved in DMSO at a concentration of 10 mM and stored at −20°C.
4
Cell Line Authentication and Maintenance
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Jurkat, MOLT4, NALM6, RS4;11, and SEM cell lines were obtained from the American Type Culture Collection (Manassas, VA), authenticated by STR profiling, and regularly tested for mycoplasma contamination (cat. no. LT07-710, Lonza, Basel, Switzerland). The TP53 isogenic NALM6 model was purchased from Horizon Discovery (Cambridge, UK). hTERT-immortalized mesenchymal stem cells (MSCs) were a gift from D. Campana (National University of Singapore, Singapore). Cells were cultured in RPMI-1640 (Sigma, St. Louis, MO) with 10% FBS and 2 mmol/L L-glutamine at 37 ℃ in humidified air supplemented with 5% CO2 and maintained in culture for no longer than 2 months and/or 25 passages.
5
Mycoplasma Detection Using MycoAlert Kit
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The MycoAlert kit was used according to the manufacturer’s instructions (Lonza, LT07-710). Briefly, 2 mL of culture media from cells before trypsinization was centrifuged at 200× g for 5 min. 100 µL of the supernatant was transferred to the well of the white 96-well plate and 100 µL of MycoAlert™ PLUS Reagent was added. The samples were incubated for 5 min at RT and the luminescence was read (Reading A, 1 s, integrated reading, Tecan Plate reader). Then, 100 µL of MycoAlert™ PLUS Substrate was added and the samples were incubated for 10 min at RT. The luminescence was read (Reading B, 1 s, integrated reading, Tecan Plate reader). The ratio of the measured values was calculated according to the formula: Reading B/Reading A. The values under 0.9 were interpreted as mycoplasma negative, values above 1.2 as mycoplasma positive. Values between 0.9 and 1.2 are a borderline. One positive control (MycoAlertTM Assay Control) and two negative controls (distilled water and culture medium) were used during testing.
6
Cell Culture Protocols for Cancer Research
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Human VCaP (ATCC® CRL2876™), HCT-116 (ATCC® CCL247™), MIA-PaCa-2 (ATCC® CRL-1420™), HCC1954 (ATCC® CRL-2338™), and 786-O (ATCC® CRL-1932™) cells were a gift from Dr. Goutham Narla at the Case Comprehensive Cancer Center, Cleveland. VCaP and HCT-116 were gifted to Hera BioLabs in April 2017. MIA-PaCa-2, HCC1954, and 786-O were gifted to Hera BioLabs in June 2018. All cell lines were originally purchased from ATCC. VCaP and MIA-PaCa-2 cells were grown in Advanced DMEM (ThermoFisher #11995065) with 10% fetal bovine serum (Atlanta Biologicals # S12450) and 1% penicillin and streptomycin solutions (Cat# 15140–122, ThemoFisher). HCT-116 cells were grown in McCoy’s 5a Medium Modified (ATCC #30–2007) supplemented with 10% fetal bovine serum (Atlanta Biologicals # S12450) and 1% penicillin and streptomycin solutions (Cat# 15140–122, Themofisher). HCC1954 and 786-O cells were grown in RPMI 1640 (ThermoFisher # A1049101) with 10% fetal bovine serum (Atlanta Biologicals # S12450) and 1% penicillin and streptomycin solutions (Cat# 15140–122, ThemoFisher). All the cells were grown in a humidified incubator at 37°C with 5% CO2. All cells lines underwent monthly testing for mycoplasma contamination (Lonza, LT07-710) and STR testing at later passages.
7
Maintenance of Human Cell Lines
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