For occlusion of the left descending coronary artery (LAD), an 8-0 Prolene suture (Ethicon, Norderstedt, Germany) was placed around it in an initial surgery and then stored subcutaneously as previously described [16 (link)]. Mice with initial surgery and 7-day recovery period without I/R were used as sham animals. After a 7-day recovery period the skin was reopened and occlusion of the LAD for 15 min with subsequent reperfusion were monitored in simultaneous ECG recording. This procedure was repeated for 1, 3, or 7 days as previously described and the hearts were excised five hours after the last ischemia episode [3 (link)]. Echocardiographic evaluation was performed in the 7-day group before heart retrieval. For histological experiments whole hearts were excised, rinsed with ice-cold cardioplegic solution, and fixated in zinc-paraformaldehyde (Z-fix, 4%; Anatech, Battle Creek, MI, USA). For mRNA studies hearts were further dissected free from great vessels and atria and immediately stored in RNA-later (Qiagen, Hilden, Germany) at 4°C [17 (link)].
8 0 prolene suture
Manufactured by Johnson & Johnson
Sourced in Germany, United States, United Kingdom
The 8-0 Prolene suture is a non-absorbable, monofilament suture material made of polypropylene. It is designed for use in fine surgical procedures, particularly in ophthalmology and microsurgery, where precise tissue approximation is required.
Automatically generated - may contain errors
Lab products found in correlation
Rnalater, by Qiagen (2 mentions)
6 0 nylon running sutures, by Johnson & Johnson (2 mentions)
Iscove modified dulbecco media (imdm), by Thermo Fisher Scientific (1 mentions)
5 0 prolene suture, by Johnson & Johnson (1 mentions)
Xylazine, by Bayer (1 mentions)
Ketamine, by Zoetis (1 mentions)
Isoflurane, by Abbvie (1 mentions)
Mouseox system, by Starr Life Sciences (1 mentions)
C57bl 6j mice, by Charles River Laboratories (1 mentions)
Pe10 tube, by BD (1 mentions)
Forene, by Abbott (1 mentions)
Osmolite 1 cal, by Abbott (1 mentions)
18 protocols using 8 0 prolene suture
1
Murine Model of Myocardial Ischemia-Reperfusion
2
Repetitive Ischemia-Reperfusion Injury Model
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Briefly, in an initial surgery, an 8-0 Prolene suture (Ethicon, Norderstedt, Germany) was placed around the left descending coronary artery and stored subcutaneously as previously described [4 (link)]. After a recovery period of 7 days following initial surgery trauma, repetitive I/R was performed. One daily episode of 15 min coronary artery occlusion was followed by reperfusion until the next day. This protocol was repeated over 1, 3, 5, or 7 consecutive days. Left ventricular function was assessed via echocardiography after 7 days I/R before heart retrieval for histology group. Hearts were excised and fixated in zinc-paraformaldehyde (Z-fix, 4%; Anatech, Battle Creek, MI, USA) for histology, stored in RNAlater (Qiagen, Hilden, Germany) for mRNA-studies, or dissolved for cardiomyocyte isolation.
3
Mouse Coronary Artery Occlusion Procedure
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The mice were anesthetized with an i.p. of 2% chloral hydrate (2 ml/100 g), endotracheally intubated by tracheotomy, and mechanically ventilated using the Inspira Advanced Safety Ventilator (ASV, NP 55–7059, Harvard Corp., Evansville, WI, USA), which supplied 0.75 ml of room air/oxygen 110 times per minute. The heart of each mouse was exposed via a left thoracotomy. After removing the pericardium, the left anterior descending coronary artery (2 mm below the tip of the left auricle) was occluded with an 8.0 Prolene suture (ETHICON, Inc., Somerville, NJ, USA). Occlusion was confirmed by observing the LV pallor immediately.
4
Intramyocardial Cell Injection in NOG Mice
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Male non-obese diabetic/severe combined immunodeficiency NOG mice (8–10 weeks old) were intubated with a 20-gauge angiocath (TERUMO, Japan) and mechanically ventilated under general anesthesia with 2% isoflurane. The heart was exposed by left anterolateral thoracotomy. Injections into the healthy hearts of NOG mice were made at a single site of the LV free wall with a total volume of 20 μl IMDM (Life Technologies, Gaithersburg, MD) supplemented with 10 μM Y-27632 containing 2.5 × 105 cells. Injection procedures were performed by a Hamilton syringe with a 30-gauge needle. For injections into damaged hearts, myocardial infarctions were generated by ligation of the left anterior descending artery using a 8-0 Prolene suture (Ethicon, Inc., Johnson and Johnson, Somerville, NJ). Myocardial infarction was confirmed by visual inspection of myocardial blanching. In treated mice, 1.0 × 106 purified iPSC-CMs were injected at 2 sites of the infarcted areas with 10 μl IMDM and 10 μM Y-27632. IMDM alone with 10 μM Y-27632 was injected into control mice.
5
Neonatal Mouse Cardiac Regeneration
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To determine the regeneration ability of neonatal mouse, we used a similar method to that used by Sadek's group.6 (link),8 (link) One‐day‐old neonatal ICR mice (n=240; Harlan Laboratories, Jerusalem, Israel) were subjected to apical resection (n=150), MI (n=90), or sham operation (n=25). To avoid pain and stress, newborn mice were anesthetized by inhalation of 2% isofluorane and 100% oxygen. The mice were cooled down on an ice bed for 4 minutes, causing asystole and reversible apnea14 (link) and preventing excessive blood loss during surgery. The cooling‐down period also provided additional anesthesia. The chest was opened by left thoracotomy, and iridectomy scissors were used to carefully and gradually resect thin segments from the left ventricle apex, as described previously.6 (link),8 (link) The thoracic wall and skin were then sutured with an 8‐0 prolene suture.
For MI induction, we repeated this anesthesia procedure and permanently occluded the mid‐LAD coronary artery by 8‐0 prolene suture (Ethicon). Finally, animals were placed on a heating pad (37°C), allowed to recover, and returned to their mothers. Sham‐operated mice underwent the same procedure except for the resection or LAD ligation. Average survival rates after apical resection and MI were 70% and 40%, respectively. Survival after sham operation was 95%.
For MI induction, we repeated this anesthesia procedure and permanently occluded the mid‐LAD coronary artery by 8‐0 prolene suture (Ethicon). Finally, animals were placed on a heating pad (37°C), allowed to recover, and returned to their mothers. Sham‐operated mice underwent the same procedure except for the resection or LAD ligation. Average survival rates after apical resection and MI were 70% and 40%, respectively. Survival after sham operation was 95%.
6
Murine Myocardial Infarction Model
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Anesthesia was induced by intraperitoneal injection (i.p.) of 100 mg/kg ketamine (Zoetis) and 10 mg/kg xylazine (Bayer Vital). Analgesia was initiated approximately 30 min before surgery by subcutaneous (s.c.) injection of 0.1 mg/kg buprenorphine. To compensate for perioperative dehydration due to blood loss and perspiration, 20 ml/kg isotonic 5% glucose solution (B. Braun) in 0.9% NaCl (9 mg/ml) was applied i.p.. Ventilation was set to a positive end-inspiratory pressure (PEEP) of 5 mbar, a respiratory rate of 110/min and an inspiration/expiration ratio of 1/1.5 with a small animal respirator (TSE Systems). Oxygen saturation, heart rate, and respiratory rate were monitored throughout the procedure by a MouseOX system (Starr Life Sciences). Anesthesia was maintained by addition of 0.5–2% isoflurane (AbbVie) during surgery. After right lateral positioning of the animal and skin disinfection, left lateral thoracotomy was performed between the 3rd and 4th rib. Opening of the pericardium allowed for identification of the left anterior descending (LAD) coronary artery. Permanent LAD ligation was performed with one single suture in the proximal middle third of the LAD, using 8-0 prolene suture (Ethicon). After evacuating the pneumothorax, chest and skin wounds were closed using a 5-0 prolene suture (Ethicon).
7
Neonatal Mouse Model of Myocardial Infarction
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Neonatal mice on P3 were anesthetized by inducing hypothermia on ice, as described previously (2 (link), 33 (link)). After transverse skin incision, lateral thoracotomy was applied at the fourth intercostal space by blunt dissection of intercostal muscles. To induce MI, an 8-0 prolene suture (Ethicon) was passed through the heart and was tied below the LAD coronary artery. Using a nonabsorbable 6-0 suture (Ethicon), the chest was stitched and skin glue was applied to join the skin together. Blood and remaining skin glue were removed with an alcohol solution. Pups were warmed under a heat lamp for several minutes until recovery and were placed back with their mother. Mice were euthanized 7 or 21 days after surgery.
8
Murine Myocardial Infarction Model
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All animal experiments were approved by the local committee on animal welfare (G-34/11, G-6/13) and were in accordance with the guidelines issued by the Federation of European Laboratory Animal Science Associations (FELASA) [16 (link)]. The healthy animal group (n = 15) consisted of C57BL/6J mice having a mean body weight of 22 ± 3 g, originally purchased from Charles River (Sulzfeld, Germany) and bred in-house for at least 8 generations. The infarct animal group (n = 5) consisted of C57BL/6J mice inflicted by ligation of the left anterior descending artery (LAD), as previously described [17 (link), 18 ]. Briefly, the surgical procedure entailed a thoracotomy between the 3rd and 4th ribs and partial removal of the pericardial sac. Following location of the LAD between the pulmonary artery and the left auricle, the LAD was ligated using an 8-0 Prolene suture (Ethicon, Norderstedt, Germany). After positioning of a chest tube between the 4th and 5th ribs, the thoracic cavity was closed in a layered manner. A postoperative period of 2 weeks was allowed before start of the imaging experiments.
9
Murine Coronary Ligation Model
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Before surgery perioperative analgesia was performed using carprofen 5 mg/kg s.c. and Temgesic 0.1 mg/kg s.c. Anesthesia was then induced with 3% isoflurane (Forene®, Abbott) and maintained with 0.8% isoflurane in 100% O2. Left parasternal thoracotomy was performed for implantation of the ligature (8-0 Prolene suture, Ethicon, Norderstedt, Germany) around the left descending coronary artery (LAD). The suture ends were threaded through a sterile PE10 tube (Becton Dickinson, Franklin Lakes, NJ, USA), exteriorized through the thoracic wall, and stored subcutaneously [30 (link)]. After chest closure, cefuroxime suspension was injected i.p. (50 mg/kg, Zinacef; Bristol-Myers Squibb, Munich, Germany) for antibiotic prophylaxis. Mice were allowed to recover for 7–10 days from initial surgery intervention.
10
Skin Grafting in Mouse Models
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Skin grafting was performed with full sterile precautions. Mice were placed prone and a 1 × 1 cm piece of skin removed from the left dorsal thorax over the costal margin. A 1 × 1 cm piece of human skin was then fashioned and its edges sutured to the mouse recipient skin with a non-absorbable 8–0 Prolene suture (Ethicon, UK). Grafts were fenestrated and covered with povidone-iodine mesh and pressure dressings which were secured with circumferential tape. Bandages were left in place for 7–10 days and then removed under general anaesthetic. Skin grafts were monitored every 1–2 days until complete loss. Graft assessment was performed independently by two researchers who were both blinded to experimental group allocations.
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