70 μm filter

Manufactured by Greiner

Sourced in Austria

The 70 μm filter is a laboratory equipment designed to separate particles or materials based on their size. It has a pore size of 70 micrometers, which allows it to filter out larger particles while allowing smaller ones to pass through. This product is intended for general filtration applications in laboratory settings.

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Lab products found in correlation

Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Collagenase type 1, by Merck Group (1 mentions) Fetal calf serum (fcs), by Merck Group (1 mentions) Collagenase 2 solution, by Worthington (1 mentions) Streptomycin, by Merck Group (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions)

3 protocols using 70 μm filter

Isolation and Culture of Primary Dermal Fibroblasts

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Primary fibroblasts were obtained from punch biopsies. The biopsies were incubated in 2.4 U/mL dispase II for 2 h at 37 °C and then separated into epidermis and dermis by hand. Dermal fibroblasts were isolated from the dermis. The dermis was incubated overnight at 37 °C with 312 U/mL collagenase type I (Merck, Darmstadt, Germany) at 37 °C. After incubation, the cell suspension was filtered through a 70 μm filter (Greiner Bio-One, Kremsmünster, Austria). The dermal fibroblasts were grown in low glucose [1 g/L] DMEM (Gibco/Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% [v/v] fetal bovine serum (FBS, (Gibco/Thermo Fisher Scientific, Waltham, MA, USA) and 1% [v/v] penicillin/streptomycin (Gibco/Thermo Fisher Scientific, Waltham, MA, USA) at 37 °C and 7% CO₂. For treatments, fibroblasts were grown in DMEM containing 2% FBS. All experiments in this study were performed on fibroblast cultures from passage 4 to 9. Working with primary cells has an impact on reproducibility. The experiments were tested multiple times before used with primary cells. Moreover, the results usually present a means of at least 3 tests per donor in order to stabilize the result.

Nickel K., Wensorra U., Wenck H., Peters N, & Genth H. (2021). Evaluation of Immunomodulatory Responses and Changed Wound Healing in Type 2 Diabetes—A Study Exploiting Dermal Fibroblasts from Diabetic and Non-Diabetic Human Donors. Cells, 10(11), 2931.

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Isolation of Murine Immune Cells

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Peritoneal lavage of mice was performed by infusing 3ml of ice cold complete RPMI media intraperitoneally and collecting the infusate. Inguinal lymph node and spleen were dissected from mice and mashed with the handle of a 3ml syringe (Falcon) against a 70μm filter (Greiner). Bone marrow was flushed with 1ml of complete RPMI media. Liver was finely minced with scissors and digested with collagenase A and bovine DNAse I in RPMI for 45 min. The liver preparation underwent two rounds of centrifugation at 30g for 3 minutes with collection of supernatant after each step. After centrifugation, the cell pellet underwent RBC lysis (Lonza).

Chow A., Schad S., Green M.D., Hellmann M.D., Allaj V., Ceglia N., Zago G., Shah N.S., Sharma S.K., Mattar M., Chan J., Rizvi H., Zhong H., Liu C., Bykov Y., Zamarin D., Shi H., Budhu S., Wohlhieter C., Uddin F., Gupta A., Khodos I., Waninger J.J., Qin A., Markowitz G.J., Mittal V., Balachandran V., Durham J.N., Le D.T., Zou W., Shah S.P., McPherson A., Panageas K., Lewis J.S., Perry J.S., de Stanchina E., Sen T., Poirier J.T., Wolchok J.D., Rudin C.M, & Merghoub T. (2021). Tim-4+ Cavity-Resident Macrophages Impair Anti-Tumor CD8+ T cell Immunity. Cancer cell, 39(7), 973-988.e9.

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Isolation and Culture of Adventitial Cells

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Upon medioadventitial splitting, the adventitial matrix was digested using collagenase II solution (Worthington Biochemical, Lakewood, NJ) according to manufacturer's instruction for a maximum of 4 hours with repeated resuspension. The suspension was strained through a 70-μm filter (Greiner Bio-One, Alphen aan den Rijn, the Netherlands) and centrifuged at 800 rpm for 5 minutes. The precipitate was resuspended in Dulbecco's Modified Eagle's Medium (Gibco by Thermo Fisher, Landsmeer, the Netherlands) supplemented with 10% fetal bovine serum (Sigma Aldrich, Zwijndrecht, the Netherlands) to inactive collagenase II activity and centrifuged at 800 rpm for another 5 minutes. After cell counting using the Countess cell counter (Invitrogen by Thermo Fisher, Landsmeer, the Netherlands) cells were seeded on plastic surfaces in 6-well plates (Costar, Sigma Aldrich) at a concentration of 10 5 cells/mL in Dulbecco's Modified Eagle's Medium supplemented with 10% fetal calf serum and 1% penicillin with streptomycin (Sigma Aldrich) as a basal medium. Mesenchymal cellular populations were selected on their ability to adhere to plastic. Cells were passaged when they reached 70% to 80% confluence.

Doderer S.A., Gäbel G., Kokje V.B.C., Northoff B.H., Holdt L.M., Hamming J.F, & Lindeman J.H.N. (2018). Adventitial adipogenic degeneration is an unidentified contributor to aortic wall weakening in the abdominal aortic aneurysm. Journal of vascular surgery, 67(6).

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