All animal experimental protocols were approved by the bioethics committee of KRIBB. For intracranial orthotopic xenograft model, single-cell suspension of control or siNNT-transfected U373MG SSEA1+ cells were prepared and injected stereotactically into the striatum of 10-week-old Balb/c-Nude mice (Dae han BioLink, Chungbuk, Korea). One million cells were injected by using Hamilton syringe (Sigma) with stereotactic device (coordinates, 2 mm anterior, 2 mm lateral, 2.5 mm depth from the dura). Six weeks after injection, the brains were perfused with 4% paraformaldehyde by cardiac perfusion and further fixed at 4 °C overnight. For subcutaneous xenograft model, single-cell suspension of 5 × 106 control, retrovial NNT-transduced or siNNT-transfected U373MG SSEA1+ cells were mixed with Matrigel (BD) and injected into the flank of 7-week-old Balb/c-Nude mice (Dae han BioLink). Tumor volume was measured using caliper daily and the tumors were excised at 6 weeks after injection and then fixed with 4% paraformaldehyde.
Balb c nude mice
Manufactured by Daehan Biolink
Balb/c nude mice are an immunodeficient mouse model characterized by the lack of a functional thymus gland, resulting in the absence of mature T cells. These mice are commonly used in biomedical research, particularly for the study of cancer, immunology, and transplantation.
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5 protocols using balb c nude mice
1
Intracranial and Subcutaneous Xenograft Models
2
Murine Xenograft Model for Cervical Cancer
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Six- to eight-week-old female Balb/c Nude mice were purchased from Daehan Biolink (Chungbuk, Korea). All animal procedures were performed under a protocol approved by the Korea University Institutional Animal Care and Use Committee (KUIACUC-2009-126). The murine fibroblast cell line NIH3T3 and human cervical cancer cell lines CUMC6, SiHa, CaSki, and HeLa were purchased from American Type Culture Collection (ATCC, Manassas, VA) and were grown in Dulbecco's Modified Eagle Medium (DMEM) in the presence of 10% fetal bovine serum (FBS). All cells were cultured in 5% CO2 balanced air at 37°C. The identities of cell lines were confirmed by short tandem repeat (STR) profiling by IDEXX Laboratories Inc. and used within 6 months for testing.
3
Triptolide Inhibits Tumor Growth in Balb/c Nude Mice
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Six-week old Balb/c nude mice were purchased from Daehan Biolink Co. (Eumseong, Korea). After 1 week acclimation, the mice were subcutaneously injected with A549 cells (5 × 106 cells/mouse) into right the dorsal flank. After 1 week, the mice were orally administered with triptolide everyday (10, 20, and 30 nmol/mouse/day). The weight of the mice and the sizes of the tumors were measured every 3 days during the experiment. The mice were sacrificed by asphyxiation with CO2, and the tumors were excised and weighed. The tumor samples were stored either for biochemical analyses or for immunohistochemistry. The animal experiment was performed under the Institutional Animal Care and Use Committee-approved Protocol (IACUC-2020-026-1) of Dongguk University (Seoul, Korea).
4
Nude Mouse Acclimation Protocol
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Six-week-old Balb/c nude mice (20–22 g, male) were purchased from Daehan Bio Link (Seoul, Korea). The mice were housed under a 12 h light/dark illumination cycle in an experimental animal room and permitted to adapt for 7 d before the commencement of the experiments. All animals received standard chow and water ad libitum, and this study was approved by the Ethical Committees for Experimental Animals of Kyung Hee University Hospital with the approval number KHMC-IACUC-20-008-01, 02.
5
Xenograft and Spontaneous Lung Tumor Models
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Six-week-old Balb/c nude mice were purchased from Daehan Biolink Co. (Eumseong, Korea) and used for xenografts. After a week acclimation, 12 mice were divided into two groups of six mice, and they were subcutaneously injected into the dorsal flank with A549 cells (5 × 106 cells/0.2 mL DMEM media) or A549-FLAG-BAP1 cells (5 × 106 cells/0.2 mL DMEM media). The body weights of mice were measured every three days during the course of study. After 18 days, mice were sacrificed by asphyxiation with CO2 and the weight of xenografts was measured.
KrasG12D/+ mice that can form lung tumors by spontaneous recombination events [16 (link)] were obtained from Dr. Tyler Jacks (Massachusetts Institute of Technology, Cambridge, MA, USA). KrasG12D/+ mice were housed in the sterile filter-capped microisolator cages and provided with water and diet ad libitum. After KrasG12D/+ mice were bred for 1 week, 8 weeks, 24 weeks, and 40 weeks, they were sacrificed, and the lung tissues were obtained. The lung tissues were stored in the deep freezer for Western blot and RT-PCR or in 10% paraformaldehyde for hematoxylin and eosin staining and immunohistochemistry.
KrasG12D/+ mice that can form lung tumors by spontaneous recombination events [16 (link)] were obtained from Dr. Tyler Jacks (Massachusetts Institute of Technology, Cambridge, MA, USA). KrasG12D/+ mice were housed in the sterile filter-capped microisolator cages and provided with water and diet ad libitum. After KrasG12D/+ mice were bred for 1 week, 8 weeks, 24 weeks, and 40 weeks, they were sacrificed, and the lung tissues were obtained. The lung tissues were stored in the deep freezer for Western blot and RT-PCR or in 10% paraformaldehyde for hematoxylin and eosin staining and immunohistochemistry.
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