The Renca cell line (syngeneic to BALB/c mice) was purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA), engineered to express firefly luciferase, and cultured as described [4 (link),29 (link),32 (link)]. Cells were confirmed negative for mycoplasma, passaged, and used at the same passage number to limit experimental variation. Intrarenal tumor challenges and bioluminescent imaging (BLI) using an IVIS Lumina III Imager (Perkin Elmer; Waltham, MA, USA; University of Alabama at Birmingham (UAB) Small Animal Imaging Facility) were performed as described [4 (link),29 (link),32 (link)]. At day six post-tumor challenge, BLI was used to assess tumor burdens in live mice prior to therapy randomization. At day seven post-tumor challenge, mice were re-injected in the tumor-bearing kidney with either sterile saline or 109 pfu of replication-deficient adenovirus (Ad) encoding a membrane-bound version of full-length murine TRAIL protein (University of Iowa Viral Vector Core) plus 100 µg CpG1826 (Integrated DNA Technologies) (AdT/CpG treatment) and further randomized to receive no therapy, InVivoMAb polyclonal Armenian hamster IgG control (BioXCell; Lebanon, NH, USA), or InVivoMAb anti (α)-mouse CTLA-4 (clone UC10-4F10-11; BioXCell) i.p. at a dose of 100 μg/mouse on days 10, 13, and 16 post-tumor challenge.
Renca cell line
Manufactured by American Type Culture Collection
Sourced in United States, China
The Renca cell line is a well-established renal carcinoma cell line derived from a spontaneously occurring renal adenocarcinoma in BALB/c mice. This cell line is commonly used in cancer research and drug development studies.
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Lab products found in correlation
Cpg 1826, by Integrated DNA Technologies (1 mentions)
Ivis lumina 3 imager, by PerkinElmer (1 mentions)
Rpmi 1640 media, by Wisent (1 mentions)
3 4 5 dimethylthiazol 2 y1 2 5 diphenyltetrazolium bromide, by Merck Group (1 mentions)
Milli q water purification equipment, by Merck Group (1 mentions)
Clear 6 well and 96 well tissue culture polystyrene plates, by Corning (1 mentions)
Sodium pyruvate solution, by Merck Group (1 mentions)
Dmem medium, by Thermo Fisher Scientific (1 mentions)
Mem non essential amino acid solution, by Merck Group (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Merck Group (1 mentions)
Streptomycin, by Merck Group (1 mentions)
L glutamine, by Merck Group (1 mentions)
Penicillin, by Sangon (1 mentions)
Streptomycin, by Sangon (1 mentions)
5 protocols using renca cell line
1
Syngeneic Renca Tumor Model with Adenoviral TRAIL
2
Subcutaneous RENCA Tumor Model
3
Renca Cell Line Maintenance Protocol
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The Renca cell line was obtained from the American Type Culture Collection Inc. (Bethesda, MD) and maintained in RPMI 1640 media (Wisent, QC) supplemented with 10% FBS (Hyclone, UT), non-essential amino acids (NEAA) (0.1 mM), sodium pyruvate (1 mM), and L-glutamine (2 mM). Cells were sub-cultivated when above 80% confluency at a 1:5 ratio. All cells were kept in a 37 °C, 5% CO2 humidified incubator.
4
Synthesis and Characterization of PEG-PLA Copolymers
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Four-armed PEG (number-average molecular weight (Mn) = 10,000 Da) was obtained from Shanghai Seebio Biotech, Inc. (Shanghai, People’s Republic of China) and used as received. DLA and LLA were provided by Changchun SinoBiomaterials Co., Ltd. (Changchun, People’s Republic of China) and recrystallized from ethyl acetate under argon atmosphere before use. Four-armed PEG-PDLA and PEG-PLLA were synthesized as our previously reported proposal [18 ]. The degree of polymerization (DP) of PLA in each arm was calculated to be 16 based on the data of proton nuclear magnetic resonance (1H NMR). The Mn of copolymer was estimated to be 14,600 g mol−1. Doxorubicin hydrochloride (DOX · HCl) was purchased from Beijing HuaFeng United Technology Co., Ltd. (Beijing, People’s Republic of China). 4′,6-Diamidino-2-phenylindole (DAPI), Alexa Fluor 488 phalloidin (Alexa 488), and 3-(4,5-dimethylthiazol-2-y1)-2,5-diphenyltetrazolium bromide (MTT) were purchased from Sigma-Aldrich (Shanghai, People’s Republic of China). RenCa cell line was obtained from the American Type Culture Collection. Clear 6-well and 96-well tissue culture polystyrene (TCP) plates were purchased from Corning Costar Co. (Cambridge, MA, USA). The deionized water used in this study was prepared through a Milli-Q water purification equipment (Millipore Co., MA, USA).
5
Establishing Stable hCAIX-Renca Cell Line
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The human HEK293T cell line and murine renal cancer Renca cell line used in the experiment were obtained from the American Type Culture Collection and Cobioer Biosciences (Nanjing, China), respectively, and passed the certificate of analysis. In a 5% CO2 atmosphere at 37°C, HEK293T cells were cultured in DMEM medium (Gibco, Invitrogen) containing 10% fetal bovine serum (FBS) (ExCell Bio), 100 U/mL penicillin (Sangon Biotech), and 100 μg/mL streptomycin (Sangon Biotech). Renca cells were supplemented with 10% FBS, 100 U/mL penicillin, 100 μg/mL streptomycin, 2 mM L-glutamine (Sigma), 1 × MEM non-essential amino acid solution (Sigma), and 1 mM sodium pyruvate solution (Sigma) in RPMI 1640 medium (Gibco, Invitrogen). All cells cultured in medium were passaged for less than 6 weeks before renewing from a frozen, early passaged population. The stable overexpressing hCAIX-Renca cells were established by Renca cells infected with CAIX lentivirus produced by HEK293T cells.
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