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Male c57b 6j mice

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Male C57B/6J mice are a commonly used inbred strain of laboratory mice. They are a widely utilized model organism in biomedical research due to their well-characterized genetic background and physiological properties.

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Vasculife vegf lifefactors kit, by Lifeline Cell Technology (2 mentions) Huvecs, by American Type Culture Collection (1 mentions)

6 protocols using male c57b 6j mice

1

Sciatic Nerve Denervation in Mice

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Male C57B/6 J mice weighing 25 g were obtained from Jackson Labs (Bar Harbor, ME) and housed in rooms with 12:12 hour light: dark cycles and controlled temperature and humidity. Animals were provided food and water ad libitum. Denervation was performed as previously described with some modifications20 (link). The animals were anesthetized by inhalation of isofluorane, hair was removed with a clipper and the skin was cleaned with betadine solution and ethanol. A small incision was made just posterior to the head of the left femur and the sciatic nerve was exposed by careful dissection. A 2 mm piece of the nerve was removed and the wounds were closed with suture and surgical glue. Animals were administered carprofen for 3 days postoperatively. Some animals received a sham nerve transection in which the nerve was exposed but not manipulated. At 7 days after nerve or sham transection, animals were anesthetized by inhalation of isofluorane and hindlimb muscles were removed by careful dissection, minimizing any pulling on the muscle. All studies with animals were approved by the IACUC at the James J. Peters VA Medical Center and were carried out in accordance with the NIH Guide for Care and Use of Laboratory Animals.
De Gasperi R., Hamidi S., Harlow L.M., Ksiezak-Reding H., Bauman W.A, & Cardozo C.P. (2017). Denervation-related alterations and biological activity of miRNAs contained in exosomes released by skeletal muscle fibers. Scientific Reports, 7, 12888.
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2

Murine HUVEC Isolation and Culture

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Male C57B/6J mice (8–12 weeks old) were purchased from Jackson Laboratory. All animal experiments were performed according to Wayne State University Institutional Animal Care and Use Committee guidelines. The primary human umbilical vein endothelial cells (HUVECs) from four different donors were obtained from the American Type Culture Collection. Cells were grown in vascular cell basal media (VCBM) supplemented with VascuLife VEGF LifeFactors Kit (Lifeline Cell Technology). Only cells between passages 2 to 5 were used for experiments.
Nguyen H., Koh J.Y., Li H., Islas‐Robles A., Meda Venkata S.P., Wang J, & Monks T.J. (2021). A novel imidazolinone metformin‐methylglyoxal metabolite promotes endothelial cell angiogenesis via the eNOS/HIF‐1α pathway. The FASEB Journal, 35(7), e21645.
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3

Impact of High-Fat Diet and Iron on Bariatric Surgery

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Male C57B6/J mice (Jackson Laboratory), age 5 weeks at arrival had ad lib access for 6 weeks to 45% high-fat regular iron diet (35ppm Fe, Dyets Inc, #115244) and water before being randomized to 4 groups based on body weight. The groups consisted of two groups receiving Sham surgery of which one group was kept on the 45% high-fat regular iron diet and the other was switched to a 45% high-fat high iron diet (350ppm Fe, Dyets Inc, #115245), the other two groups received VSG surgery (details are described below) of which one group was kept on the 45% high-fat regular iron diet and the other was switched to 45% high-fat regular iron diet. Designated high iron diet groups were switched to the diet 2 weeks prior to surgery. At 56 days post-surgery an ip-gtt (2g/kg glucose) was performed. Animals were terminated at day 70 post surgery using CO2 and cardiac puncture. Animals were not fasted before termination. At termination, blood was collected to determine hematocrit, total iron levels, and hepcidin levels. Duodenum and ileum were dissected and processed for mRNA expression determination (detailed description below).
Evers S.S., Shao Y., Ramakrishnan S.K., Shin J.H., Bozadjieva-Kramer N., Irmler M., Stemmer K., Sandoval D.A., Shah Y.M, & Seeley R.J. (2022). Gut HIF2α signaling is increased after VSG, and gut activation of HIF2α decreases weight, improves glucose, and increases GLP-1 secretion. Cell reports, 38(3), 110270.
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4

Male C57/B6J Mice Protocol

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Male C57/B6J mice aged 12–14 weeks (Jackson Labs) were housed in groups of 4 (12-hr light/dark cycle, ambient temperature of 22 ± 3°C, with food and water available ad libitum). All animal procedures and experimental designs were approved by the Veterans Affairs Palo Alto Health Care System Institutional Animal Care and Use Committee and followed the “animal subjects” guidelines of the International Association for the Study of Pain.
Tajerian M., Hung V., Khan H., Lahey L.J., Sun Y., Birklein F., Krämer H.H., Robinson W.H., Kingery W.S, & Clark J.D. (2016). Identification of KRT16 as a Target of an Autoantibody Response in Complex Regional Pain Syndrome. Experimental neurology, 287(Pt 1), 14-20.
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5

Mouse Endothelial Cell Culture

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Male C57B/6J mice (8‐12 weeks old) were purchased from Jackson Laboratory. All animal experiments were performed according to Wayne State University Institutional Animal Care and Use Committee guidelines. The primary human umbilical vein endothelial cells (HUVECs) from four different donors were obtained from the American Type Culture Collection. Cells were grown in vascular cell basal media (VCBM) supplemented with VascuLife VEGF LifeFactors Kit (Lifeline Cell Technology). Only cells between passages 2 to 5 were used for experiments.
Nguyen H., Koh J.Y., Li H., Islas‐Robles A., Meda Venkata S.P., Wang J, & Monks T.J. (2021). A novel imidazolinone metformin‐methylglyoxal metabolite promotes endothelial cell angiogenesis via the eNOS/HIF‐1α pathway. The FASEB Journal, 35(7), e21645.
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6

Impact of High-Fat Diet and Iron on Bariatric Surgery

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Male C57B6/J mice (Jackson Laboratory), age 5 weeks at arrival had ad lib access for 6 weeks to 45% high-fat regular iron diet (35ppm Fe, Dyets Inc, #115244) and water before being randomized to 4 groups based on body weight. The groups consisted of two groups receiving Sham surgery of which one group was kept on the 45% high-fat regular iron diet and the other was switched to a 45% high-fat high iron diet (350ppm Fe, Dyets Inc, #115245), the other two groups received VSG surgery (details are described below) of which one group was kept on the 45% high-fat regular iron diet and the other was switched to 45% high-fat regular iron diet. Designated high iron diet groups were switched to the diet 2 weeks prior to surgery. At 56 days post-surgery an ip-gtt (2g/kg glucose) was performed. Animals were terminated at day 70 post surgery using CO2 and cardiac puncture. Animals were not fasted before termination. At termination, blood was collected to determine hematocrit, total iron levels, and hepcidin levels. Duodenum and ileum were dissected and processed for mRNA expression determination (detailed description below).
Evers S.S., Shao Y., Ramakrishnan S.K., Shin J.H., Bozadjieva-Kramer N., Irmler M., Stemmer K., Sandoval D.A., Shah Y.M, & Seeley R.J. (2022). Gut HIF2α signaling is increased after VSG, and gut activation of HIF2α decreases weight, improves glucose, and increases GLP-1 secretion. Cell reports, 38(3), 110270.
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