Bs 70 xbs

Manufactured by Biosharp

Sourced in China

The BS-70-XBS is a high-precision balance designed for laboratory use. It features a weighing capacity of up to 70 grams and a readability of 0.1 milligrams. The balance is equipped with a backlit LCD display for easy reading of measurements.

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10 protocols using bs 70 xbs

Thymocyte Phenotypic Analysis by Flow Cytometry

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The thymus tissues obtained from 2.5.2 were ground and sieved through 70 µm filters (BS-70-XBS; Biosharp, Hefei, China). FITC Hamster Anti-Mouse CD3e (BD553061), APC Rat Anti-Mouse CD4 (BD553051), PE Rat Anti-Mouse CD8a (BD553032), and BB700 Hamster Anti-Mouse TCRβ (BD745846) were adopted to stain isolated thymocytes. The flow cytometry (CytoFlex S, Beckman) collected all flow data and analyzed the result using CytExpert 4.1.
Collect thymocytes obtained from 4.6. FITC Hamster Anti-Mouse CD3e (BD553061), APC Rat Anti-Mouse CD4 (BD553051), PE Rat Anti-Mouse CD8a (BD553032) and BB700 Hamster Anti-Mouse TCRβ (BD745846) were adopted to stain thymocytes. The flow cytometry (CytoFlex S; Beckman, Brea, CA, USA) collected all flow data and analyzed the result using CytExpert 4.1.

Ying Y., Tao N., Zhang F., Wen X., Zhou M, & Gao J. (2024). Thymosin β4 Regulates the Differentiation of Thymocytes by Controlling the Cytoskeletal Rearrangement and Mitochondrial Transfer of Thymus Epithelial Cells. International Journal of Molecular Sciences, 25(2), 1088.

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Isolation of Primary Bone Marrow Macrophages

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Mice were sacrificed and immersed in 75% alcohol for 5 min. We separated the tibia and femur of the limbs under sterile conditions, cut both ends of the bone, and washed the bone cavity repeatedly with Dulbecco's Modified Eagle Medium (DMEM, Cat#C11995500BT, Gibco, USA) until it was transparent. We filtered the cells with a 70-µm cell strainer (Cat#BS-70-XBS, Biosharp, Hefei, China). Red blood cells were lysed and removed. After centrifugation, the cells were collected and cultured in DMEM containing 40 μg/mL recombinant mouse M-CSF (Cat#315-02, PeproTech, Suzhou, China). We replaced half of the complete culture medium three days later with fresh DMEM containing M-CSF. We completely replaced the culture medium on Day 5 and continued culturing for 1-2 days to obtain primary bone marrow macrophages for further study.

Chen Y., Wang T., Liang F., Han J., Lou Z., Yu Y., Li J., Zhan T., Gu Y., Dong L., Jiang B., Zhang W., Wu M, & Lu Y. (2024). Nicotinamide phosphoribosyltransferase prompts bleomycin-induced pulmonary fibrosis by driving macrophage M2 polarization in mice. Theranostics, 14(7), 2794-2815.

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Isolation of Decidua Cell Populations

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Human tissues of the decidua including two compartments were collected immediately after delivery (37 (link)). In brief, the decidua basalis and parietalis were collected from basal plate and chorioamniotic membranes, respectively. Tissues were washed with ice-cold sterile phosphate buffer saline (PBS) to remove the blood and then divided into two parts (Figure 1). One part was fixed with polyformaldehyde, embedded in paraffin and cut into sections (3 μm) for immunohistochemical analysis, while the other part of fresh decidua basalis and parietalis (20 g) were rinsed thoroughly with sterile PBS containing 1% penicillin/streptomycin (Cat. 10378016, Gibco) to remove the blood, and were then cut and homogenized using a gentleMACS Dissociator (Miltenyi Biotec) in RPMI 1640 medium supplemented with 0.25% Trypsin (Cat. G4001, Servicebio), collagenase Type IV (Cat. G5027, Servicebio) at 1.0 mg/ml concentration, and deoxyribonuclease I (Cat. B002004, Sangon Biotech) at 0.1 mg/ml concentration (15 (link), 37 (link)). Subsequently, homogenized tissues were digested for 60 min at 37°C with gentle agitation, then filtered through a 70 μm cell strainer (Cat. BS-70-XBS, Biosharp) and washed in PBS to obtain single-cell suspensions, which were stored in liquid nitrogen for future flow cytometry analysis.

Zha Y., Liu H., Lin X., Yu L., Gao P., Li Y., Wu M., Gong X., Bian X., Kang Q., Zhi P., Dang X., Wang J., Feng L., Qiao F., Huang Y, & Zeng W. (2022). Immune Deviation in the Decidua During Term and Preterm Labor. Frontiers in Immunology, 13, 877314.

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Isolation of Choroidal Endothelial Cells and Pericytes

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Primary choroidal endothelial cells (CECs) and pericytes were isolated from C57BL/6 mice (4–6 weeks old) as shown in the previous studies^{11 (link),12 (link)}. Briefly, choroidal tissues were digested with 0.05% trypsin (25300120, Gibco, USA) at 37 °C for 30 min. After removing RPE layers, they were digested in 5 ml of digestion solution containing Collagenase II (200 U/ml, 2275MG100, BioFroxx, China) and DNase I (30 U/mL, D7073, Biosharp, China) at 37 °C for 1 h. Then, the cell mixtures were filtered using a 70-μm filter (BS-70-XBS, Biosharp, China). CECs were isolated using anti-CD31 antibody-coated Dynabeads (Invitrogen, USA). The freshly isolated CECs were cultured on collagen IV (17104019, Gibco, USA) coated 6-well plates in the Microvascular Endothelial Growth Medium (EGM2-MV; CC-3202, Lonza) at 37 °C in a humidified atmosphere of 5% CO₂. Primary pericytes were isolated using anti-PDGFRβ antibody-coated Dynabeads (Invitrogen, USA) and cultured in DMEM with 10% FBS at 37 °C and 5% CO₂. CECs and pericytes within the three passages were used for experiments.

Shen Y., Xu M., Ren L., Li X., Han X., Cao X., Yao J, & Yan B. (2023). A novel retinoic acid drug, EYE-502, inhibits choroidal neovascularization by targeting endothelial cells and pericytes. Scientific Reports, 13, 10439.

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Isolation of Ear Skin Cells

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Animals were euthanized 48 h after intravenous injection of MSCs. The ear was removed at the level of the base, and the dorsal and ventral skin was peeled off and placed in 2 mL digestion buffer. After incubation for 1 h at 37 °C, the ear skin was pestled into a 70 µm cell strainer (Biosharp, BS-70-XBS). Strainers were washed with 2 mL 1% FBS and 2 mM EDTA in PBS. The cells were then treated with 1×RBC lysis buffer (Beyotime, C3702) for 2 min at 4 °C, washed with PBS and analyzed using a BD/FACS Celesta flow cytometer.

Ye T., Liu X., Zhong X., Yan R, & Shi P. (2023). Nongenetic surface engineering of mesenchymal stromal cells with polyvalent antibodies to enhance targeting efficiency. Nature Communications, 14, 5806.

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Gut Organoid Culture Protocol

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Gut organoid cultures were prepared according to reported methods (24 (link)). Briefly, crypts were incubated for 15 min at room temperature in gentle cell dissociation reagent (STEMCELL Technologies). Then, crypts were passed through a 70-μm cell strainer (Biosharp Life Sciences, BS-70-XBS) and collected by centrifugation at 100 × g for 5 min. The crypts were embedded with Matrigel (Corning) and plated, after which the plate was carefully transferred to a 37°C incubator and incubated at 37°C for 10 min to allow domes to polymerization. After polymerization of Matrigel, the crypts were added into IntestiCult medium (STEMCELL Technologies) and the plate was placed in an incubator at 37°C and 5% carbon dioxide. The culture medium was fully exchanged three times per week.

Wang Y., Gao Y., Su X., Hao Y., Zhang Y, & Yang R. (2024). LNCGM1082 in Gut Epithelial Cells Promotes Expulsion of Infected Epithelial Cells and Release of IL-18. ImmunoHorizons, 8(1), 35-46.

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Decellularized Porcine Intestinal ECM

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Each batch of dECM was fabricated from three piglets' submucosal tissue. A conjoint decellularization strategy modified from established protocols for porcine intestine was used subsequently. To initiate decellularization, submucosal tissue pieces were placed in pure water (Spring‐R10; RSJ, Xiamen, China) overnight at 4°C. Washed tissue was decellularized in 4% SDC (30970; Sigma‐Aldrich, St. louis, MO, USA) for 4 h at room temperature. After SDC treatment, the tissue was washed with pure water for 24 h at 4°C, with water change every 6 h. Afterwards, a step of 2000kU DNase‐I (11284932001; Sigma‐Aldrich) in 1 M NaCl was performed for 3 h at room temperature. The tissue was washed with pure water again for 48 h at 4°C, with water changed every 6 h. A rotator at 60 rpm (DS‐S 100; Servicebio, Wuhan, China) was used throughout the decellularization and rinsing. Rinsed tissue was lyophilized (LGJ‐12; HUAYU XIONGDI, Zhengzhou, China) for 72 h and milled (Tissuelyser‐24; Jingxin, Shanghai. China) into a fine powder that could pass through a 70 μm cell strainer (BS‐70‐XBS; Biosharp, Anhui, China). Acquired dECM powder was stored at −20°C until further use.

Xu Z., Huang J., Liu Y., Chen C., Qu G., Wang G., Zhao Y., Wu X, & Ren J. (2022). Extracellular matrix bioink boosts stemness and facilitates transplantation of intestinal organoids as a biosafe Matrigel alternative. Bioengineering & Translational Medicine, 8(1), e10327.

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Isolation and Differentiation of Mouse Bone Marrow-Derived Macrophages

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The femora and tibiae from C57BL/6J were flushed with 5ml ice cold RPMI-1640 medium to isolate mouse bone marrow. Bone marrow cells were filtered by a 70µm cell strainer (BS-70-XBS, Biosharp) and red blood cell lysis buffer (R1010, Solarbio) was utilized to lyse red blood cells. The remaining cells were cultured in RPMI-1640 medium supplemented with 20ng/ml of mouse recombinant macrophage-colony stimulating factor (315–02, Peprotech, NJ, USA) for 7 days. The culture medium was changed every 2 days to obtain BMDMs for further studies.

Mao J., Chen Y., Zong Q., Liu C., Xie J., Wang Y., Fisher D., Hien N.T., Pronyuk K., Musabaev E., Li Y., Zhao L, & Dang Y. (2024). Corilagin alleviates atherosclerosis by inhibiting NLRP3 inflammasome activation via the Olfr2 signaling pathway in vitro and in vivo. Frontiers in Immunology, 15, 1364161.

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Single-cell Ovarian Immune Profiling

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The single-cell suspension was prepared using mechanical dissociation. Briefly, the isolated ovaries were thoroughly washed with PBS, and the fatty tissue was removed. The ovaries were then placed on a 70 μm cell filter (Biosharp, BS-70-XBS), and the tissue was ground with the rubber head of a 5 ml syringe to obtain a single-cell suspension. The prepared single-cell suspension was washed once with PBS and stained with the following monoclonal antibodies for cell surface phenotypic analysis: CD45-efluor 450 (eBioscience, 48-0451-82), CD11b-APC cy7 (eBioscience, 47-0112-82), F4/80-PE (eBioscience, 12-4801-82), CD86-FITC (eBioscience, 11-0862-82); CD206-APC (eBioscience, 17-2061-82), and IL-6R-PE (eBioscience, 12-1261-80). For IL-6R, rat IgG2b kappa isotype control (eB149/10H5) and PE (eBioscience, 12-4031-82) were used as isotype controls. The single-cell suspension was stained with antibodies for 30 min at 4 °C in the dark, and single-color controls were also prepared for compensation adjustment. The samples were washed and resuspended in PBS. Flow cytometry data acquisition was performed using Cytoflex LX (Beckman Coulter, Brea, CA). Data analysis was conducted using FlowJo 10.0. Supplementary Fig. 3 provides an example of the gating strategy.

Hu B., Zheng X, & Zhang W. (2024). Resveratrol-βcd inhibited premature ovarian insufficiency progression by regulating granulosa cell autophagy. Journal of Ovarian Research, 17, 18.

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Isolation and Culture of Primary Chondrocytes

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The cartilage was cut into small pieces about 1–2 mm in diameter and then washed with phosphate-buffered saline (PBS, Gibco Life Technologies, Grand Island, NY, USA) more than 2 times. Then, the cartilage pieces were added to Dulbecco’s modified Eagle’s medium/Ham’s F-12 nutrient mixture (DMEM/F12, Gibco Life Technologies, USA), containing 1% penicillin (100 IU/mL) and 1% streptomycin (100 μg/mL), and 5% fetal bovine serum (FBS, Gibco Life Technologies), with additional protease (4 mg/mL medium), at 37 °C for 1.5–2 h. Next, the solution was poured off, and the same medium solution as above was added, except that the protease was replaced by collagenase P (0.25 mg/mL medium), at 37 °C for 6–8 h. Finally, primary chondrocytes were collected and seeded in T75 flasks after filtration using a 70 µm cell strainer (BS-70-XBS; Biosharp, Hefei, China) and centrifugation at 1200 rpm for 3 min. All primary chondrocytes were cultured in a DMEM/F-12 medium containing 5% FBS and 1% penicillin/streptomycin solution.

Liu H., Deng Z., Yu B., Liu H., Yang Z., Zeng A, & Fu M. (2022). Identification of SLC3A2 as a Potential Therapeutic Target of Osteoarthritis Involved in Ferroptosis by Integrating Bioinformatics, Clinical Factors and Experiments. Cells, 11(21), 3430.

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