In the example we describe, mice were treated with 0.4 mg of drug on days −2 and−1 prior to bone marrow transplantation (day 0). Lung damage was analyzed 20, 30, 40, and 50 weeks after irradiation by computed tomography scans and histology and compared with a sham-irradiated, age-matched, control group. The irradiation procedure, itself, is painless. Embryonic Stem (ES) Cell Culture/Mouse Embryonic ... Then 4 hours later the mice are injected (via tail vein) with bone marrow cells. Much of the seminal work regarding mouse strain and age sensitivity was done more than 50 years ago. While local irradiation alone could control the primary tumor in a dose-dependent manner, it did not improve mouse survival. Senescence in Mouse Hippocampus Next we established a 3-cycle irradiation protocol, in which mice were sub-lethally irradiated once monthly over a 3 month period. The easiest and most commonly used method experimentally is total-body irradiation (TBI), which is achieved by placing the mice in specifically designed irradiators; the dose of whole-body gamma irradiation causes the animals to become either transiently or chronically immunosuppressed. NSG ™ Variants Portfolio Mice were bred under isolated conditions and maintained in filter-top, micro-isolator cages. For mouse irradiation, a dose rate of about 10 Gy/min (Bragg peak) was applied to deliver physical doses (“high”, “intermediate”, “low”) of 80, 60, and 40 Gy to C3H/He mice, and of 85, 65, and 45 Gy to C57BL/6 mice. Evaluation of Acute Reactions on Mouse Skin Irradiated ... Activation of p53 After Irradiation Impairs the ... D. Making drinking water readily available. • Our irradiation protocol is 1000 rads (we use a 137Cesium Gammacell source) for a C57BL6/J mouse. 12. Bone Marrow Transplantation in Mice Other studies used either body molds with no fixation points for the attachment of the mouse head or glue to fixate collimators on mouse skin [14,33,37]. The . Engrafts a wide range of solid and hematological cancers. Production of Hematopoietic Chimeras Care of Irradiated Mice A. (a) Diagram illustrating the device and experimental protocol used for the generation of a mouse model of UV irradiation‒induced itch. Mice in the control and TBI groups were given PBS in the same protocol. 09-1042 and No. Cancer patients undergoing cranial irradiation are at risk of developing neurocognitive impairments. Mice in the control group showed a gradual increase in body weight and significantly higher increase ratio than those in the IR group (IR group: 2.96±6.47%; control group: 15.5±4.80%; n=11 each, P<0.001; Figure 2A). IRRADIATION Preparation of Mouse Embryonic Fibroblast ... - CSH Protocols Food and acidified tap water were provided ad libitum. Optimal protocol for total body irradiation for allogeneic bone marrow transplantation in mice. CF1 MEFs are traditional feeder cells for culturing pluripotent stem cells; for MEFs derived from ICR mice, see Mouse (ICR) Inactivated Embryonic Fibroblasts . For acute lymphoblastic leukemia (ALL), the 5-year survival rate has improved significantly since 1975. Mouse Visualization of the sperm could be accomplished by Hoechst staining and ultraviolet (UV) light irradiation. The previous studies have suggested that irradiation at the early postnatal days of mice induced the impairment of hippocampal neurogenesis [8, 10, 11], but the long-term effect of irradiation at different postnatal days on the structure of the hippocampus and brain remains unknown. 09-1042 and No. Identify gene type (for instance ACR+, ACR‐) by genotyping. The camera (FSV-210L, Apiste) of the thermograph was fixed so that the long axis of … 2.4. The study paradigm of the experi-ments is shown in Table 1. It should be noted that the protocols included in this manual are intended to serve as a guide only, and optimization of culture protocols is encouraged to ensure success. Antibiotic treatment – Prior to irradiation animals will be treated with For mouse irradiation, a dose rate of about 10 Gy/min (Bragg peak) was applied to deliver physical doses (“high”, “intermediate”, “low”) of 80, 60, and 40 Gy to C3H/He mice, and of 85, 65, and 45 Gy to C57BL/6 mice. To determine the effects of the clinical dosage administration in combination with approved pharmaceutical drugs on the survival of mice exposed to high-dose TBI (7 Gy), 4 combinations of G-CSF and RP were administered to mice within 2 h after X … Irradiation is repeated 3 hrs later for a total dose of 700R -1200R (split dose irradiation is used to limit the non-hematopoietic toxicity -- usually intestinal damage). In the 7.5Gy irradiation experiment, 10 mice wereusedineachgroup, whileinthe4Gyirradiationexperi-ment,5micepergroup.10daysafter4GyTBI,themicewere sacrificed and samples were collected [13]. Choose recipient C57BL/6 mice that are 6-10 weeks old. In effect, with an imaging system inside the irradiation cabinet, it is possible: C. Use of antibiotics in the drinking water. 1. Wean and keep male mice at 21‐day age. Lethal irradiation of C57/B6 mice: 950-1050 Rads administered in a split dose (3 hrs. H&E staining of mouse skin 8 hours post pulse laser irradiation in the absence and presence of Au-NRs. In this study, the effect of ATX on hematopoietic system injury after TBI was investigated. irradiation, mice were euthanized by CO 2 inhalation followed by cervical dislocation. IRRADIATION PROTOCOL The following is what we do using the Dartmouth Irradiation Facility for bone marrow chimerism using either pure C57Bl/6, C57Bl/6.SJL, or Rag-/-γC-/-animals (C57Bl/10XC57Bl/6). After activation, oocytes were incubated in Hoechst 33342 or 33258 stain and exposed to UV irradiation. The miRNA microarray data revealed that a total of 18 miRNAs were differentially expressed (>2-fold; P<0.05) in LLC1 cells exposed to all irradiation protocols . RAD SOURCE Technolgies RS2000 is an excellent device to accomplish this. • Inject a known number of leukemia cells from a donor mouse into a recipient mouse to determine the number of cells needed to cause leukemia. We describe a method using targeted peripheral irradiation to induce fatigue-like behavior in mice. 1A). We asked whether irradiation induced senescence in mouse hippocampus, and whether p53 or p21 played a role in this response. Doses or irradiation protocols other than these will be noted as an exception to the standard Mice in the IR group showed a reduced body weight at 2 weeks after irradiation, which recovered by 4 weeks. However, it also promotes accelerated senescence in healthy tissues and leads to progressive cognitive dysfunction in up to 50% of tumor patients surviving long term after treatment, due to γ-irradiation-induced … 10 days after 4 Gy TBI, the mice were sacrificed and samples were collected [13]. Sensitive to irradiation. Injecting tumor cells in rodents, usually mice, is an accepted experimental procedure for the purpose of either propagating a tumor line or for studying various cancers and cancer treatments. b. 2. In the neutron irradiation study of a mouse PCNSL model, the therapeutic effect of BNCT on PCNSL was evaluated in terms of survival. In the 7.5 Gy irradiation experiment, 10 mice were used in each group, while in the 4 Gy irradiation experiment, 5 mice per group. Beam arrangement for multi-mouse proton irradiation. 09-1049-1, No. Mouse Surgery Protocols (edited 1/11/96) A. Gamma-irradiation will be administered at the dose rate of 400 Rads/minute using 137 Cs source located in Room 112 of Paige laboratory. Mice in the control and TBI groups were given PBS in the same protocol. To induce apoptosis, 0.35 μg of mouse recombinant TNF-α (amino acids 80 to 235, R&D Systems) in a volume of 50 μl was injected into the tail vein of mice, 24 hours before irradiation. Irradiation preferentially kills rapidly dividing cells, including bone marrow and epithelial cells of the gut and other organs. 4. As a tool for studying the mechanisms of hippocampal-dependent cognitive decline, we developed a mouse model replicating the results of the recent clinical RTOG 0933 Made from 3.2mm ( 1 / 8 ) clear acrylic, the mouse irradiation chamber features a flip-top lid for easy loading and unloadingReusable animal cages constructed from durable materials are designed to house and breed of small animals. However, (n = 4 mice/group). Left panels: Histological features (hematoxylin and eosin) following exposure on hairless mice skin for each wavelength with threshold limits. mouse model protocol without irradiation. Irradiation . 1. Choose recipient C57BL/6 mice that are 6-10 weeks old. 2. The night before irradiation, remove their food but not their water. 3. Next morning, put 3 mice per clean Tupperware container - 1 in each division. Wipe down each container with 70% EtOH, and then add some bedding to the containers so that the mice are comfortable. Mouse B16F1 melanoma (3), MCA/129 fibrosarcoma (4), and human HCT116 C57BL/6 mice were placed into six treatment groups and received the following irradiation treatments: 600 MeV/n 56 Fe (0.2 Gy), 1 GeV/n 16 O (0.2 Gy), 350 MeV/n 28 Si (0.2 Gy), 137 Cs (1.0 Gy) gamma rays, 137 Cs (3.0 Gy) gamma rays, and sham irradiation. In contrast to irradiated wild type (WT) mice, TNF-a deficient (TNF-a KO) mice or WT mice treated with a TNF-a-neutralizing antibody were partially protected from the apoptotic effects of irradiation. An exception to a Guideline must be described and justified in the Animal Protocol and approved during the normal review process. Mice were anesthetized by intraperitoneal injection of 0.3% pentobarbital (20 ml/kg). Irradiation of mice with ionizing radiation has been shown to induce a lasting increase in IL1B concentration in peripheral blood. The night before irradiation, remove their food but not their water. One of the possible mechanisms may be demethylation of CpG cytosines in the Il1b promoter, which has not been characterized in detail for the mouse. 2. Currently, feeder cells are mitotically inactivated either by gamma irradiation 24,25,26,27,28,29,30 or MMC 3,4,11,31,32,33,34. Considerations. Irradiation and humanization induced changes in white blood cells and platelets counts of NSG mice. 3. In contrast to irradiated wild type (WT) mice, TNF-a deficient (TNF-a KO) mice or WT mice treated with a TNF-a-neutralizing antibody were partially protected from the apoptotic effects of irradiation. The bodybuilders use Clenbuterol in at main cycles. This is the same strain, sex and age of mice that was used in our previous studies on functional and histological changes after kidney irradiation (4-6). (A) Comparison of peripheral white cell count (WBC) and differentials (neutrophils, Neu; lymphocytes, Ly; monocytes, Mo) between young 6 month old (YM), middle age 12 month old (MM) and humanized (HM) NSG mice. Following whole-brain irradiation, polymerase chain reaction (PCR) arrays for senescence-associated genes showed increased expression of CDKN1A ( p21 ) and CDKN2A ( p19ARF ) in mouse hippocampus at 9 weeks. 2.3 Irradiation treatment of mice Our previous study showed that a dose of LD50 at day 30 after ray exposure to C57BL/6 mice was 6 Gy [21]. ... the cells were fixed with methanol and acetic acide based on a protocol used in our previous publication (Fotouhi et al., 2015). Compromised human stem cell regeneration. Mouse Holder for Microbeam Irradiation 2.4. Mice were placed in the left lateral decubitus position and the dorsal skin was irradiated using the laser system. Next morning, put 3 mice per clean Tupperware container - 1 in each division. Nos.GSC-6001G and GSC-6201G, these widely-used MEF cells are isolated from outbred CF1 mice, mitotically arrested by irradiation, and tested to ensure safety and performance. Irradiation can provoke the translocation of bacteria from the gastrointestinal tract … Histological evaluations of effects on mice skin after UV irradiation. USE OF THE RPO IRRADIATION ROOM Mouse Strain Irradiation Protocol References - As a guide, complete myeloablation is likely to be achieved in any immunocompetent (adult) strain at, (1) UQ Animal Ethics Committee - Standard Operating Procedure . Total whole body doses will not exceed 500 Rad. Preparation of mitotically inactivated feeder cell layer (treatment with mitomycin C or γ-irradiation) is also described. W e studied, and successfully eradicated, a mouse population on a small. Recently, cabinet-size, X-ray-based irradiators have been advertised as a … Meanwhile, treat the mouse with neomycin containing acid water for the first 2–3 weeks after irradiation. The protocols provided in this unit describe aspects of isolation and expansion of MEFs and maintenance of established feeder cells. into recipient mouse, determine TD50. protocol described (2), employing a revised PA2 primer (5′-GGCTACCCGTGATATTGC-3′), and 35 cycles of PCR amplification, each at 94°C for 15 sec, 64°C for 30 sec, and 68°C for 90 sec. H. Rodents may or may not be anesthetized for irradiation. Made from 3.2mm ( 1 / 8 ) clear acrylic, the mouse irradiation chamber features a flip-top lid for easy loading and unloadingReusable animal cages constructed from durable materials are designed to house and breed of small animals. In contrast to irradiated wild type (WT) mice, TNF-α deficient (TNF-α KO) mice or WT mice treated with a TNF-α-neutralizing antibody were partially protected from the apoptotic effects of irradiation. G. Housing. Laser Irradiation Protocol. We have previously demonstrated, using chimeric resistant MRL/lpr mice, that a … 17-2006) involving mice mentioned above and descripted in the irradiation section were reviewed and approved by The The comparative parameter most frequently referenced is the LD50/30, which is the dose of whole body irradiation that is lethal to 50% of a target population by 30 days after exposure. ESCs are pluripotent, meaning that they are able to differentiate into all derivatives of the three primary germ layers (ectoderm, endoderm, and mesoderm). In the adult mouse brain, the SVZ contains a heterogeneous population of proliferating cells. and 100 cells of each type scored for each mouse for the number of grains per labeled cell. Five mice were utilized for each point with 500 cells of each type scored for each mouse for the labeling index (L.I.) In this study, we investigated dose, dose rate and time-dependent effects in a dose range of 0.005 to 2 Gy on immune parameters after whole body irradiation (IR) using a pro-inflammatory (ApoE−/−) and a wild … Animal preparation for irradiation: a. Acidified water – Animals to be irradiated will be placed on acidified water (pH 2.5-3.0) from 1 week prior to 2 weeks post irradiation. To exclude negative effects of this treatment, we examined toxicity on activated mouse oocytes. The hematopoietic system is especially sensitive to total body irradiation (TBI), and myelosuppression is one of the major effects of TBI. Mouse Surgery Protocols (edited 1/11/96) A. Gamma-irradiation will be administered at the dose rate of 400 Rads/minute using 137 Cs source located in Room 112 of Paige laboratory. Commonly, irradiators produce or may be set to deliver about 85 - 165 cGy/min. Feed the irradiated mouse with sterilized CD for recovery of 30 days ad libitum. Embryonic stem cells (ESCs) are derived from the inner cell mass of day 5-6 blastocysts. Single exposure - Mice will be irradiated with 800-950 rads (exact dose is strain dependent) at a rate of approximately 80 rads/minute. This dose rate is achieved when the irradiator is used with out attenuators in turntable position 3 for a period of 10-12 minutes. mice. Upper panel: the UV-emitting device (1) targeting a small skin area is placed on the metal shelf (2) and parallel to The several different, complex protocols are enumerated in Results (Table I). The cell culture protocol described here includes the in vitro culture of mouse ES cells in serum-containing media using mouse embryonic fibroblasts and ESGRO mLIF. Irradiation . Send DDC to Research Diets, Inc for preparing OpenStandard Diet with added compound DDC 0.1% after gamma-Irradiation. All mice 41 and 43 days old at the start of irradiation. Unirradiated mice were also given injections of either saline or SR 4233. Left liver lobes were collected at 30, 60, 120, 270, and 360 days post-irradiation. Total whole body doses will not exceed 500 Rad. F. Provision of softened food. In the C57BL6/J mouse liver, irradiation induced p53 phosphorylation (Ser15), stabilized p53, and induced p21 transcription but did not induce transcription of puma, another downstream target of p53 (Fig. Rats, mice, hamsters, gerbils, and guinea pigs will be adequately contained and remain safe during transportation. Rats, mice, hamsters, gerbils, and guinea pigs will be adequately contained and remain safe during transportation. Rodent Irradiation and Bone Marrow Transplant Procedures . This study investigated the therapeutic effects of combined local irradiation and anti-HER2/neu antibody in a mixed tumor mouse model comprised of a nonmetastatic neu-positive tumor and a metastatic neu-negative tumor. Irradiation typically renders mice immunosuppressed; depending on dosage, this effect may be transient (lasting until hematopoietic cells are replenished through endogenous recovery or transplant) or permanent. During this period, mice are susceptible to opportunistic infections that may affect survival or model performance. After the irradiation, remove the lead piece from the mice and put them to the home cage. Importantly, the activity of the NRF2 gene, an important regulator of the antioxidant cellular machinery, was completely suppressed by irradiation but was restored by post-irradiation Prdx6 treatment. Finally, we conducted the IFA to study the e®ect of irradiation on senile plaques in mouse brains. Protocol : 1. X-irradiation. Analysis of the Numbers of Bone Marrow Mononuclear Cells (BMMNCs), HPCs, and … Protocol II: Mr PB lineage tracing by parabiosis In the WBR group, the mouse was placed in a 1×1 cm 2 irradiation range from the post-canthus line to the post-aurem line. ab1796 staining Histone H3 in mouse liver tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Irradiation Activated p53 and Suppressed Liver Regeneration in C57BL6/J Mice. Summary: Mouse Embryonic Fibroblast (MEF) cells are required to support the growth of undifferentiated mouse or human ES cells, iPS cells, thus they are also called feeder cells.MEF cells are isolated from mouse embryos and are used at their early passages. Survival rate following the clinical dosage administration in combination with approved pharmaceutical drugs. A new irradiation plan was established on the Small Animal Radiation Research Platform machine for local heart irradiation of mice with single doses of 8 and 16 Gy. Next we established a 3-cycle irradiation protocol, in which mice were sub-lethally irradiated once monthly over a 3 month period. Recent evidence suggests that radiation-induced injury to the hippocampi could play an important role in this cognitive decline. • Irradiate donor mouse, collect and count leukemic cells, inject i.p. Immune system problems at gvhd mouse protocol without irradiation as eosinophils and contains supplemental material for these defects are sincerely grateful to gvhd mouse model protocol without irradiation as you. 09-1049-1, No. Flow cytometry was used to detect the proportion of hematopoietic … IRRADIATION PROTOCOL The following is what we do using the Dartmouth Irradiation Facility for bone marrow chimerism using either pure C57Bl/6, C57Bl/6.SJL, or Rag-/-γC-/-animals (C57Bl/10XC57Bl/6). Mice will be monitored to assure there is no acute illness. No thymic lymphomas, can be used for long & short-term experiments. BB-UVB irradiation evokes scratching behavior in mice. non-radiation, immunocompetent mouse strains [2–4] and on the effects of irradiation of stem cells before engraftment [5]. Andras Nagy, Marina Gertsenstein, Kristina Vintersten, and Richard Behringer; Cold Spring Harb Protoc; 2006; doi: 10.1101/pdb.prot4400 Each dose group contained three animals; one sham-irradiated animal per strain served as control. Greater tolerance to genotoxic agents compared to NSG. To induce apoptosis, 0.35 μg of mouse recombinant TNF-α (amino acids 80 to 235, R&D Systems) in a volume of 50 μl was injected into the tail vein of mice, 24 hours before irradiation. A copy of one of the most relevant reports accompanies A new experimental topical treatment protocol was assessed in mice receiving orthovoltage X-rays at an equivalent dose to that applied to human breast cancer patients in … The selected non-lethal irradiation dose leads to a week-long reduction in voluntary wheel-running activity. In order to achieve a total dose of 40 mg/kg of busulfan, 0.8 mg of busulfan will need to be administered to a 20 gram mouse. Lethal irradiation of C57/B6 mice: 950-1050 Rads administered in a split dose (3 hrs. Recipient mice are given 850-1100 r whole body lethal irradiation from an x-ray or gamma irradiation source. Euthanasia (Guideline) Guidelines: The IACUC has provided a set of guidance documents (Policies, Guidelines, and Informational Sheets) for use when planning animal procedures at the University of Iowa. C3H mice were irradiated with (5+5) or (6+6) Gy at intervals of 1 to 24 h. A single-dose irradiation (12 Gy) was also given to some mice. BALB/c BMCs (3 × 10 6) were injected intravenously into the irradiated mice 24 h after the last irradiation. Astaxanthin (ATX) is a powerful natural anti-oxidant with low toxicity. The irradiation procedure, itself, is painless. Isocenter is set at the center of the restraint system such that with a spot-scanning proton system, the incident beams on top and bottom shelf positions are divergent. Tissue was fixed with paraformaldehyde, permeabilized with 0.05% Triton X-100 in PBS for 30 minutes and blocked with 5% BSA for 1 hour; antigen retrieval was by heat mediation in sodium citrate pH 6. This is given in one dose. español (Spanish) svenska (Swedish) Türkçe (Turkish) March 17th, 2017. Before MEF cells are used as feeder cells, they must be treated by irradiation or mitomycin C to stop the cells from further dividing. All experimental protocols (Experimental Animal Research Plan No. 3. • TD50 = number of cells needed to cause leukemia in 50% of recipient animals. • For mice that will be fed an atherogenic diet, start the diet 6 weeks after transplantation Anti-inflammatory low-dose therapy is well established, whereas the immunomodulatory impact of doses below 0.1 Gy is much less clear. Lung damage was assessed in irradiated and sham-irradiated animals by Annexin-Vivo 750 imaging (NEV-11052, PerkinElmer). In mouse models of bone marrow transplantation, irradiation of recipient animals to ablate endogenous hematopoietic tissues is conventionally given by radioisotope-based irradiators. However, there is currently very limited research comparing the effects of using either Cs-137 or X-ray irradiation of immune-deficient mice for the pur-pose of performing stem cell transplants. To investigate whether the abscopal effects of cranial irradiation (C-irradiation) cause testicular damage in mice, male C57BL/6 mice (9weeks of age) were randomly divided into a sham irradiation group, a shielded group and a C-irradiation group and administered sham/shielded irradiation or C-irradiation at a dose rate of 2.33Gy/min (5Gy/d for 4 d consecutively). Read Article. The mice were between eating or drinking (e.g., difficulty with ambulation). A colorwash map shows the final composite plan from two lateral opposed beams.
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