An osmotic pump (ALZET Micro-Osmotic Pump model 1002; DURECT) filled with 1.5 mg/kg body weight of tacrolimus (Prograf; Astellas Pharma) was subcutaneously implanted under the back skin of the BALB/c mice (n = 71) to create an immunosuppressed allogeneic model. As one pump can continuously deliver the solution for 14 days, another new pump was added every 13 days to ensure constant release of tacrolimus. Plasma concentrations of tacrolimus were assessed by electrochemiluminescence immunoassay.
Alzet micro osmotic pump model 1002
Manufactured by Durect
Sourced in United States
The ALZET Micro-Osmotic Pump model 1002 is a laboratory equipment product designed for continuous and controlled delivery of test substances. It is a self-contained, osmotically-driven pump that operates without the need for programming or external power sources. The pump delivers the test substance at a constant rate over a pre-determined period of time, ranging from 1 to 14 days.
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4 protocols using alzet micro osmotic pump model 1002
1
Tacrolimus-Induced Immunosuppression in BALB/c Mice
2
Hind Limb Ischemia in Aged Mice
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The mice received a separate mini-pump for co-infusion of MitoTEMPO (Santa Cruz Biotechnology, Dallas, TX, USA) or saline. The mice were anesthetized by pentobarbital (50 mg/kg body weight, i.p.) and an osmotic mini-pump (ALZET micro-osmotic pump, model 1002, DURECT Co., Cupertino, CA, USA) was subcutaneously implanted. MitoTEMPO (180 µg/kg/day) and saline for the old mice and saline for the young mice were continuously infused. From the previous study concerning to the hemodynamic effect of MitoTEMPO, we determined 180 µg/kg/day of MitoTEMPO is a suitable dose [15 (link),49 (link)]. Unilateral hind limb ischemia was induced by ligation of the left femoral artery from its origin just below the inguinal ligament under tribromoethanol anesthesia (200 mg/kg body weight, i.p.) at day 7 after pump implantation [51 (link)]. The sham operation was without femoral artery ligation but with skin incision in the right hind limb of ischemia-induced mice. The mice were sacrificed with isoflurane aspiration and a lethal dose of pentobarbital (60 mg/kg body weight i.p.) on POD 2 or 21, and examinations were performed. We selected POD 2 as early phase of ischemia, because the expressions of HIF-1α and VEGF were more remarkable than other days in our preliminary test.
3
Cardiac effects of MitoTEMPO in mice
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The mice received a separate minipump for co-infusion of MitoTEMPO or saline. The mice were anesthetized by pentobarbital (50 mg per body, intraperitoneally) and an osmotic minipump (ALZET micro-osmotic pump, model 1002; DURECT Co., Cupertino, California, USA) was subcutaneously implanted. MitoTEMPO (180 µg/kg/day) and saline as a control for the old mice and saline for the young mice were infused continuously. Blood pressure was monitored using the tail-cuff method. Previously, we measured the systolic blood pressure and heart rate in mice administered MitoTEMPO (50, 180, and 500 µg/kg/day) with an osmotic minipump at 14 days after infusion. When treated with 500 µg/kg/day of MitoTEMPO, systolic blood pressure decreased over 5% (P<0.05) compared with before treatment, which are identical to the results of a previous report 7 (link). Left ventricular diastolic function (E/e′) and heart rate did not change with or without administration of 50 µg/kg/day of MitoTEMPO; that is, the effect of MitoTEMPO is doubtful as the dose is too small. Therefore, we considered the suitable dose of MitoTEMPO to be 180 µg/kg/day. Twenty-eight days after surgery, the animals were killed with a lethal dose of pentobarbital and examinations of the coronary artery, cardiac myocytes, and aorta were performed.
4
Subcutaneous Tacrolimus Osmotic Pump
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An osmotic pump (ALZET Micro-Osmotic Pump model 1002; DURECT Corporation, Cupertino, CA, USA) filled with 1.5 mg/kg body weight of tacrolimus was subcutaneously implanted under the abdominal skin of the BALB/c mice to create an allogeneic model of immunosuppression. As one pump can continuously deliver the solution for 14 days, another new pump was added every 13 days to ensure the constant release of tacrolimus.
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