Teratomas were generated in severe combined immunodeficient (SCID) mice from 201B7 hiPSCs grown under ESF-Fb-EzF conditions for more than 10 passages. The cells harvested by dispase were resuspended in DMEM supplemented with RI (10 μM). The cells from a confluent single well in a 6-well plate were injected into the thigh muscle of a SCID (C.B-17/lcr-scid/scidJcl) mouse (CLEA Japan, Tokyo, Japan). Nine weeks after injection, tumors were dissected, weighed, and then fixed with 10% formaldehyde Neutral Buffer Solution (Nacalai Tesque, Kyoto, Japan). Paraffin-embedded tissue was sectioned and stained with hematoxylin and eosin (HE). All animal experiments were conducted in accordance with the guidelines for animal experiments of the National Institute of Biomedical Innovation, Osaka, Japan.
C b 17 lcr scid scidjcl
Manufactured by CLEA Japan
Sourced in Japan
The C.B-17/lcr-scid/scidJcl is a laboratory mouse strain that is commonly used in research applications. It is characterized by a severe combined immunodeficiency (SCID) phenotype, which results in a lack of functional T and B cells. The strain is also deficient in the recombinase-activating gene (RAG), leading to an absence of mature lymphocytes. This mouse model is often utilized for the study of immune system function and development, as well as for the evaluation of cell-based therapies and xenotransplantation experiments.
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6 protocols using c b 17 lcr scid scidjcl
1
Teratoma Generation from hiPSCs in SCID Mice
2
Islet Cell Sheet Transplantation in Diabetic SCID Mice
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All animal experiments were performed with the approval of the Institutional Animal Care and Use Committee of Tokyo Women's Medical University. Male Lewis rats, 8–12 weeks of age (LEW/CrlCrlj, Charles River, Yokohama, Japan) were used as donors for islet cell sheet transplantation. Male severe combined immunodeficient (SCID) mice, 7–10 weeks of age (C.B-17/lcr-scid/scidJcl, CLEA Japan, Tokyo) were used as recipients. Diabetes was induced in the SCID mice by a single intraperitoneal injection of streptozocin (220 mg/kg body weight). Non-fasting blood glucose levels were measured in blood samples collected from the tail vein of each mouse with the use of a handheld blood glucose meter (Glutest Neo Super; Sanwa Chemistry Laboratory, Nagoya, Japan). SCID mice with hyperglycemia greater than a non-fasting blood glucose level of 350 mg/dL were used for the transplantation of islet cell sheets. All animals were kept under a controlled environment (22–24 °C, 55 ± 10% humidity, and a 12-h light/dark cycle).
3
Teratoma Formation Assay in Mice
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The cells were harvested by dispase treatment, collected into tubes, and centrifuged, and the pellets were suspended in DMEM. A total of 1,000,000 hESCs were injected into the rear leg muscle or thigh muscle of a SCID (C.B-17/lcr-scid/scidJcl) mouse (CLEA Japan, Tokyo, Japan). Seven weeks after injection, tumors were dissected, weighed, and fixed with 10% formaldehyde Neutral Buffer Solution (Nacalai tesque, Kyoto, Japan). Paraffin-embedded tissue was sliced and stained with hematoxylin and eosin. All animal experiments were conducted in accordance with the guidelines for animal experiments of the National Institute of Biomedical Innovation, Health, and Nutrition, Osaka, Japan.
4
SCID Mouse Model for Immunology
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Seven-week-old male severe combined immune- deficient (SCID) mice (C.B-17/lcr-scid/scid Jcl) were provided by CLEA Japan, Inc. (Tokyo, Japan). All animals were housed in a specific pathogen-free environment under controlled conditions (temperature, 20–26°C; humidity, 30–70%; light/dark cycle, 12/12 h) and were allowed to acclimatize and recover from shipping-related stress for more than 5 days prior to the study. Chlorinated water and irradiated food were provided ad libitum. The health of the mice was monitored by daily observation. The humane endpoints were deterioration of general conditions and sacrifice in the event of a body weight loss exceeding 20%. All animal experiments were performed at Chugai Pharmaceutical Co., Ltd. The experiments were reviewed and approved by the Chugai Pharmaceutical Co., Ltd., Institutional Animal Care and Use Committee.
5
Teratoma Formation Assay for Human iPSCs
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For teratoma formation assay, human iPS cells that dissociated into single cells after long-term culture in our culture system were injected directly into the subrenal capsular space of immunodeficient mice (C.B-17/lcr-scid/scidJcl; CLEA Japan, Tokyo, Japan). After 3 months, the teratomas were surgically dissected out of the mice and fixed with 4% paraformaldehyde solution (Wako Pure Chemical Industries). For immunocytochemistry, each fixed teratoma was sectioned into 7-μm slices using a cryostat. All animal experiments in this study were approved in advance by the University of Tokyo and were conducted in accordance with rules of its Institutional Animal Care and Use Committee.
6
SCID Mice for Cell Transplantation
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For this study, male 8-week-old SCID mice (CB-17/lcr-scid/ scidJcl) were purchased from CLEA Japan, Inc (Tokyo, Japan). C57BL/6-Tg(ubiquitinÀgreen fluorescent protein [GFP]) 30scha/J mice were purchased from The Jackson Laboratory (Bar Harbor, ME, USA) and used for cell transplantation. Mice were housed under a 12-h light/dark cycle and allowed free access to food and tap water.
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