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Lignin in Biological Systems
Published in Severian Dumitriu, Valentin Popa, Polymeric Biomaterials, 2020
Lignin-derived macromolecules (LDMs) that are highly sulfonated (LSA) are negatively charged compounds that exhibit a number of biological activities. LSA and other LDMs are potent inhibitors of HIV in vitro, possibly through interference with the CD-4 receptor/HIV interaction. In vitro microbicidal activity of LDMs has been demonstrated with several other viruses as well. This antiviral and antipathogen activity is cell surface mediated and is likely related to similar inhibitory effects of other sulfonated and sulfated macromolecules. Selective effects of LDMs on various cell types have also been observed. For example, an LDM has been shown to dramatically inhibit the growth of fibroblast and sarcoma cells, but yet activate murine macrophages and induce proliferation of bone marrow cells. LDMs have also been shown to inhibit fertilization in echinoderms without showing cytotoxic effects through a mechanism that involves inhibition of the sperm acrosome reaction. LSA inhibits the sea urchin sperm acrosome reaction and it competes with the natural sulfated ligand, egg jelly, on the sperm surface. LSA binds to the head of capacitated macaque sperm, a location consistent with its biological activity. LSA significantly inhibited the binding of macaque sperm to macaque zonae pellucidae both when the compound was added to sperm after capacitation and when it was added to sperm before Percoll separation and capacitation (Tollner et al. 2002).
Zearalenone perturbs the circadian clock and inhibits testosterone synthesis in mouse Leydig cells
Published in Journal of Toxicology and Environmental Health, Part A, 2021
Lijia Zhao, Yaoyao Xiao, Cuimei Li, Jing Zhang, Yaojia Zhang, Meina Wu, Tiantian Ma, Luda Yang, Xiaoyu Wang, Haizhen Jiang, Qian Li, Hongcong Zhao, Yiqun Wang, Aihua Wang, Yaping Jin, Huatao Chen
LCs were isolated from 2–3-month-old adult PER2::LUC knock-in mice or WT mice and cultured as previously described (Chen et al. 2017; Lin et al. 2015). Briefly, testes were de-capsulated and digested using 0.05% collagenase I in DMEM/F12 (Invitrogen, Carlsbad, CA) for 15 min at 34°C. The suspended LCs were collected by filtration using a 0.074 mm nylon cell strainer, and purified over a discontinuous Percoll gradient (5%, 30%, 58%, and 70%). After centrifugation at 800 × g at 4°C for 30 min, the majority of the purified LCs was found to be concentrated in the third layer. The LCs were seeded on 35 mm collagen-coated dishes in 2 ml DMEM/F12 containing 10% fetal bovine serum (FBS) supplemented with antibiotics and cultured in a humidified atmosphere of 5% CO2 at 37°C for 2 days until reaching 70–80% confluence. TM3 mouse cells (The Cell Bank of Type Culture Collection of Chinese Academy of Sciences, Shanghai, China) were seeded on 35 mm collagen-coated dishes and cultured in DMEM (Gibco, Grand Island, NY, USA) supplemented with 2.5% FBS, 5% horse serum and 1 × antibiotic-antimycotic (AA, containing penicillin, streptomycin, and amphotericin B; Invitrogen) in a humidified atmosphere of 5% CO2 at 37°C. All cells were incubated with various concentrations of ZEA for the indicated times.
The growth and pluripotency of mesenchymal stem cell on the biodegradable polyurethane synthesized with ferric catalyst
Published in Journal of Biomaterials Science, Polymer Edition, 2018
Qianqian Wei, Jiachang Jin, Xinyuan Wang, Qijun Shen, Mi Zhou, Shizhong Bu, Yabin Zhu
Bone marrow (BM) was harvested from vertebra of rabbit (The animal center of Ningbo University, China). First, general anesthesia via ear vein inject 3% nembutal, shaving the fur, antisepsis was done with 75% alcohol, then incision of skin and exposure spine, a puncture needle was introduced into spine for bone narrow cell harvesting. After collection, 5 ml percoll solution, which density is 17.2 g/cm3, it was prepared by percoll, 8.5% NaCl and 0.85% NaCl, was put into centrifuge tube, then slowly added 5 ml bone marrow cell harvesting, spin for 5 min at 2500 r/min. Mononulcear fraction (WF) was isolated in a percoll gradient. The WF was washed once and cultured in RPMI-1640, supplemented with 10% fetal bovine serum.
Two modified density gradient centrifugation methods facilitate the isolation of mouse Leydig cells
Published in Preparative Biochemistry & Biotechnology, 2023
Jiayang Jiang, Xiaoman Zhou, Chunliu Gao, Rongqin Ke, Qiwei Guo
As shown in Figure 1b, 1 mL of 100% Percoll solution was premixed with 1 mL of crude cell solution. The mixture was used as a 50% Percoll solution to prepare the discontinuous Percoll gradients (30, 40, 50, 60, 70, and 80% from bottom to top, with 2 mL in each layer). The gradient solution was centrifuged at 1800 × g for 20 min at 4 °C. Approximately 0.5 mL of cell solution was collected from the interface between 50 and 60% Percoll based on the visible cell layer. The washing process was identical to that of the regular method. The experiments were performed in triplicate.