Mice were administered a senolytic cocktail containing 5 mg/kg dasatinib (D‐3307, LC Labs) and 50 mg/kg quercetin (Q4951, Sigma‐Aldrich) as described by Xu et al. (Xu et al., 2018 (link)). Briefly, 7.5 mg of dasatinib and 75 mg of quercetin were dissolved in 5 ml of 10% polyethylene glycol 400 (PEG 400; 202398, Sigma‐Aldrich). We chose this volume of PEG 400 because it allowed us to gavage a 30–45 gram mouse with 100–150 μl of D+Q, which is less than the approximate stomach volume of 400 μl in adult mice (McConnell et al., 2008 (link)). Mice were gavaged bi‐weekly for 4 months (Figure 1a ) with D+Q (n = 20 young and n = 19 old) or vehicle (n = 20 young and n = 19 old; 10% PEG 400) using 20‐gauge disposable polypropylene feeding tubes (FTP‐20‐30, Instech, Plymouth Meeting, PA). These mice were then divided into 3 groups of n = 6–7/group.
Ftp 20 30
Manufactured by Instech
Sourced in United States
The FTP-20–30 is a laboratory equipment designed for fluid transfer and processing. It has a flow rate range of 20-30 liters per minute. The core function of the FTP-20–30 is to facilitate the movement and handling of liquid or fluid samples within a controlled laboratory environment.
Automatically generated - may contain errors
Lab products found in correlation
Dasatinib, by Merck Group (2 mentions)
Quercetin, by Merck Group (2 mentions)
Peg400, by Merck Group (2 mentions)
3 mm aortic punch, by Medtronic (2 mentions)
Q4951, by Merck Group (1 mentions)
Male sprague dawley rat, by Charles River Laboratories (1 mentions)
Mouse diet 5015, by Purina (1 mentions)
C57bl 6j mice, by Jackson ImmunoResearch (1 mentions)
7 protocols using ftp 20 30
1
Senolytic Cocktail Administration in Mice
2
Senolytic Cocktail Administration in Mice
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Mice were administered a senolytic cocktail containing 5 mg/kg dasatinib (D-3307, LC Labs, Woburn, MA) and 50 mg/kg quercetin (Q4951, Sigma-Aldrich, St. Louis, MO) in a similar manner as described by Xu et al. [21 (link)] and consistent with a previous study by our laboratory [9 ]. Briefly, 7.5 mg of dasatinib and 75 mg of quercetin were dissolved in 5 mL of 10% polyethylene glycol 400 (PEG 400; 202398, Sigma-Aldrich). We chose this volume of PEG 400 because it allowed us to gavage a 30–45 g mouse with 100–150 μL of senolytic, which is much less than the approximate stomach volume of 400 μL in adult mice [36 (link)]. Senolytics or vehicle (10% PEG) was administered on day 7 and day 10 of the 14-day mechanical overload protocol (Fig. 5a ) using 20-gauge disposable polypropylene feeding tubes (FTP-20–30, Instech, Plymouth Meeting, PA). We chose a hit-and-run approach based on the work from the Kirkland laboratory and these time points because we previously showed that SA β-Gal + cells appear as early as 7 days in a muscle injury model [9 ]. Furthermore, MOV places a constant stress on the muscle such that senescent cells would continue to appear after the initial D + Q gavage and it is unlikely that D + Q kills all of the senescent cells after a single treatment.
3
Cannulation Techniques for Cardiac Preservation
4
Rat Kidney Perfusion and Preservation
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All animal experiments were approved by the Institutional Animal Care and Use Committee (IACUC) at the University of Minnesota (IACUC Protocol: 1905‐37029A). Male Sprague‐Dawley rats (Charles River Laboratories, Wilmington, MA) weighing 176–300 g were used in this study. General anesthesia was induced by 4% isoflurane and maintained with 1.5% of isoflurane and 1 liter‐per‐minute oxygen. Details on surgery are described in Supporting Information. Briefly, the infrarenal abdominal aorta was cannulated using a 20G bulb tip catheter (FTP‐20‐30, Instech Laboratories, Plymouth Meeting, PA) and secured using a 4‐0 silk ligature. A second catheter (male luer, 45518‐46, Cole Parmer, Vernon Hills, IL) was placed in the inferior vena cava for venous drainage. The suprarenal aorta and vena cava were ligated using silk ligatures and 10 mL University of Wisconsin (UW) solution containing 500 IU of heparin was perfused at 0–4 °C. The kidney was mobilized from the Gerota's fascia and kept in UW at 4 °C for further study.
5
Housing and Feeding of C57BL/6J Mice
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C57BL/6J mice were purchased from The Jackson Laboratory (Bar Harbor, ME, United States). All the mice were housed, four mice per cage, upon arrival and were acclimated on standard chow (Purina Mouse Diet 5015, 25.34% fat, 19.81% protein, 54.86% CHO, 3.7 kcal/g; Lab Diet, St. Louis, MO, United States) for one week. They had free access to water, unless otherwise noted, and maintained on a 12-h light and 12-h dark cycle with lights on from 0500 to 1700 h. Oral gavage was performed using single-use, sterile plastic feeding tubes (20 ga × 30 mm; cat# FTP-20-30, Instech Laboratories, Plymouth Meeting, PA, United States). The animal care protocol was approved by the Institutional Animal Care and Use Committee of Rutgers University (OLAW #A3262-01); Protocol #999900014.
6
Cannulation Techniques for Cardiac Preservation
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This cannulation approach was performed in the same manner as the above-written method for type A cannulation, but following that procedure an 18G needle was used to puncture the LV and a 20G, 5–8 mm long bulb tip catheter (FTP-20–30, Instech laboratories) was inserted into the LV. Afterward, 5 mL of UW and 125 IU of heparin were injected through the cannula inside the ascending aorta and the heart was placed in a container of cold UW solution.
Other tested cannulation methods include the following: Type A cannulation with 20G bulb tip catheter insertion into the LV via the left atrial appendage puncture, and after successful completion of type A cannulation, a small defect was created on the left atrial appendage and the mitral valve of the heart was removed using a 3 mm aortic punch (Medtronic, USA).
Other tested cannulation methods include the following: Type A cannulation with 20G bulb tip catheter insertion into the LV via the left atrial appendage puncture, and after successful completion of type A cannulation, a small defect was created on the left atrial appendage and the mitral valve of the heart was removed using a 3 mm aortic punch (Medtronic, USA).
7
Rat Kidney Isolation and Perfusion
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Rats were anesthetized with isoflurane, and a cruciate laparotomy was performed. The bowel and mesentery were retracted. The abdominal aorta, inferior vena cava, and left renal artery and vein were mobilized for cannulation. The kidney was dissected free of Gerota’s fascia. A 20G bulb tip catheter (FTP-20-30, Instech Laboratories, Plymouth Meeting, PA) was used to cannulate the abdominal aorta below the renal arteries. A second cannula (Male Luer to Hose Barb Adapter, 45518-46, Cole Parmer, Vernon Hills, IL) was used to cannulate the inferior vena cava for venous drainage. A third cannula (PE-10-100 polyethylene tubing; 0.011″ ID × 0.025″ OD; SAI Infusion Technologies) was used to cannulate the ureter. The suprarenal aorta was cross clamped, and 15 mL of University of Wisconsin (UW) solution containing 500 IU of heparin was perfused at 0–4 °C. The suprarenal aorta and vena cava were ligated and divided above the left renal vessels. The kidney was explanted and stored in UW at 4 °C until use (30 min to 2 h).
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