SKRC-52 cells were grown to 80% confluence and detached with Trypsin-EDTA 0.05% (Life Technologies). Cells were washed with Hank’s Balanced Salt Solution (HBSS, pH 7.4) once, counted and re-suspended in HBSS to a final concentration of 3.4 × 107 cells/ml. Aliquots of 5 × 106 cells (150 µl of a suspension) were injected subcutaneously in the right flank of female athymic Balb/c nu/nu mice (6-8 weeks of age, Janvier).
Balb c nu nu mice
Manufactured by Janvier Labs
Sourced in France
BALB/c nu/nu mice are an athymic mouse strain, characterized by a lack of functional T cells due to a spontaneous mutation in the Foxn1 gene. These mice are commonly used in research as a model for studying immune function and as a host for human tumor xenografts.
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Lab products found in correlation
Trypsin edta 0.05, by Thermo Fisher Scientific (3 mentions)
Prism 6, by GraphPad (2 mentions)
Superfrost plus, by Thermo Fisher Scientific (1 mentions)
Matrigel, by Avantor (1 mentions)
Shandon cryotome fse, by Thermo Fisher Scientific (1 mentions)
C57bl 6 mice, by Janvier Labs (1 mentions)
Hanks balanced salt solution (hbss), by Thermo Fisher Scientific (1 mentions)
Hanks balanced salt solution (hbss), by Janvier Labs (1 mentions)
Roswell park memorial institute (rpmi), by Thermo Fisher Scientific (1 mentions)
Cobra γ counter, by Hewlett-Packard (1 mentions)
15 protocols using balb c nu nu mice
1
Xenograft Tumor Engraftment in Athymic Mice
2
Xenograft Tumor Growth Inhibition Assay
3
Xenograft Model for Head and Neck Cancer Research
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The patient-derived
head and neck squamous
cell carcinoma (HNSCC) xenograft model SCCNij15333 (link) was used for in vivo experiments. Six
to eight week old female athymic BALB/c nu/nu mice (Janvier Laboratories,
Le Genes-Saint-Ile, France) were implanted subcutaneously with a tumor
(2 mm diameter) on the right hind leg. Animals were included when
tumors were >80 mm3 (4–5 weeks after implantation).
Allocation to the treatment groups was block-randomized by tumor size.
The studies were approved by the Central Authority for Scientific
Procedures on Animals (RU-DEC-2015-0071) and carried out under the
supervision of the local Animal Welfare Body. Note that ethical approval
for the mice experiments in this study was provided on September 8,
2015 by the institutional Animal Welfare Committee of the Radboud
University Medical Center (application no. AVD103002015209), in accordance
with the guidelines of the Revised Dutch Act on animal experimentation.
head and neck squamous
cell carcinoma (HNSCC) xenograft model SCCNij15333 (link) was used for in vivo experiments. Six
to eight week old female athymic BALB/c nu/nu mice (Janvier Laboratories,
Le Genes-Saint-Ile, France) were implanted subcutaneously with a tumor
(2 mm diameter) on the right hind leg. Animals were included when
tumors were >80 mm3 (4–5 weeks after implantation).
Allocation to the treatment groups was block-randomized by tumor size.
The studies were approved by the Central Authority for Scientific
Procedures on Animals (RU-DEC-2015-0071) and carried out under the
supervision of the local Animal Welfare Body. Note that ethical approval
for the mice experiments in this study was provided on September 8,
2015 by the institutional Animal Welfare Committee of the Radboud
University Medical Center (application no. AVD103002015209), in accordance
with the guidelines of the Revised Dutch Act on animal experimentation.
4
Dose Escalation in Athymic Mice
5
Subcutaneous Tumor Xenograft Model
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3 × 106 SK-RC-52 cells in 100 µL PBS were injected subcutaneously into the right flank of 6–8 week old male BALB/c nu/nu mice (Janvier, le Genest-Saint-Isle, FR). Two different dimensions (length and width) of tumor size were measured at least twice a week with a caliper. The tumor volume was calculated using the equation: V = (π/6)*(higher diameter)*(lower diameter)2. All animal experiments were performed in accordance with German law and guidelines for care and use of laboratory animals. The experiments were approved by the competent authority (Landesuntersuchungsamt Rheinland-Pfalz, Germany; according to §8 Abs. 1 Tierschutzgesetz; permission no. 23177-07/G15-1-033).
6
Xenograft Tumor Growth Inhibition Assay
7
Subcutaneous Xenograft Model with SKRC-52 Cells
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SKRC-52 cells
were grown as described above to 80% confluence and detached with
trypsin–EDTA 0.05% (Life Technologies). Cells were rinsed once
with Hank’s balanced salt solution (HBSS, pH 7.4), and counted
and suspended again in HBSS to give a final concentration of 3.4 ×
107 cells/mL. Aliquots of 5 × 106 cells
(150 μL of the suspension) were injected subcutaneously into
the right flank of athymic BALB/c nu/nu mice (8–10 weeks old
females, Janvier).
were grown as described above to 80% confluence and detached with
trypsin–EDTA 0.05% (Life Technologies). Cells were rinsed once
with Hank’s balanced salt solution (HBSS, pH 7.4), and counted
and suspended again in HBSS to give a final concentration of 3.4 ×
107 cells/mL. Aliquots of 5 × 106 cells
(150 μL of the suspension) were injected subcutaneously into
the right flank of athymic BALB/c nu/nu mice (8–10 weeks old
females, Janvier).
8
Xenograft Tumor Induction and Tissue Harvesting
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10×106 LNCaP, SKOV-3, or PC3 cells per mouse in 50% Matrigel (VWR, Radnor, U.S.A.) were subcutaneously inoculated into the right shoulder of 7-8-week-old BALB/c nu/nu mice (body mass 20-25 g) obtained from Janvier (La Genest-Saint Isle, France). The tumours were allowed to grow for 4-5 weeks until they reached ~0.5-0.75 cm3 in size as determined using a caliper. 5×105 B16.F10 cells per mouse were implanted subcutaneously in the right dorsal flank of 8-week-old C57BL/6 mice (body mass 20-25 g) purchased from Janvier (La Genest-Saint Isle, France). The tumours were allowed to grow for 10-15 days until they reached ~0.5-0.75 cm3 in size as determined using a caliper. Subsequently, the mice were anesthetized with 2.5% isoflurane in O2 at a flow rate of 1 L/min after which they were sacrificed by decapitation. Tumour or muscle tissue was excised, rinsed with saline to remove blood and rapidly frozen in 2-methylbutane (-40 °C). Next, 20 µm sections were obtained using a cryotome (Shandon cryotome FSE; Thermo Fisher, Waltham, MA) and these were mounted on adhesive microscope slides (Superfrost Plus; Thermo Fisher Scientific) and stored at -20 °C.
9
Sunitinib Treatment on RCC Xenografts
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Institutional guidelines were strictly followed for maintenance of animals and experimental procedures were approved by the Institutional Animal Care and Use Committee (IACUC). Female BALB/c nu/nu mice, 6 to 8 weeks of age, were obtained from Janvier, France, and maintained at the local central animal facility. Animals were either grafted s.c. with freshly excised NU12 RCC xenograft pieces [24] (link) of approximately 1 to 2 mm3 or injected s.c. with 2*106 freshly trypsinized SK-RC-52 cells. Treatment with sunitinib (SU11248, Sutent®, Pfizer, NY) was started when tumors had reached a tumor volume of 100 to 200 mm3.
Sunitinib was dissolved in 0.1 M Na-citrate, pH 4.5 and freshly prepared every week.
Mice received the equivalent of 40 to 50 mg/kg (0.8 to 1 mg/200 μl) sunitinib or vehicle orally per day for 7 to 14 days.
Sunitinib was dissolved in 0.1 M Na-citrate, pH 4.5 and freshly prepared every week.
Mice received the equivalent of 40 to 50 mg/kg (0.8 to 1 mg/200 μl) sunitinib or vehicle orally per day for 7 to 14 days.
10
Subcutaneous Xenograft Establishment
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SK-RC-52 cells were
grown to 80–100% confluence and detached with Trypsin–EDTA
0.05% (Invitrogen). Cells were washed once with Hank’s Balanced
Salt Solution (HBSS, Thermo Fisher Scientific, pH 7.4), counted, and
resuspended in HBSS. Aliquots of 5–10 × 106 cells were resuspended in 150 μL of HBSS and injected subcutaneously
in the right flank of female athymic BALB/c nu/nu mice (8–10
weeks of age, Janvier).
grown to 80–100% confluence and detached with Trypsin–EDTA
0.05% (Invitrogen). Cells were washed once with Hank’s Balanced
Salt Solution (HBSS, Thermo Fisher Scientific, pH 7.4), counted, and
resuspended in HBSS. Aliquots of 5–10 × 106 cells were resuspended in 150 μL of HBSS and injected subcutaneously
in the right flank of female athymic BALB/c nu/nu mice (8–10
weeks of age, Janvier).
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