KUSA-A1 mouse osteoblastic cell line was obtained from JCRB Cell Bank (Japanese Collection of Research Bioresources Cell Bank; JCRB1119). Primary periodontal ligament cells were isolated and cloned as described previously (41). Cells were maintained in α-minimum essential media (Wako Pure Chemical Industries, Japan) containing 10% fetal bovine serum (Thermo Fisher Scientific, MA, USA) and kanamycin (60 μg/ml, Wako Pure Chemical Industries) under normal cell culture conditions (37°C, 5% CO2). For osteogenic differentiation, the cells were cultured in the same media supplemented with l-ascorbic acid (50 μg/ml) (013-19641, Wako Pure Chemical Industries) and 10 mM β-glycerophosphate (193-02041, Wako Pure Chemical Industries). Calcein [C001, Dojindo, Japan; stock solution (1 mg/ml) was prepared by dilution from stock (50 mg/ml) in 1 M KOH] was incubated at 1 μg/ml for Ca2+ imaging during culture with normal or osteogenic media. For mineral staining, cells were fixed with cold 100% ethanol for 5 min and stained with 1.0% Alizarin Red S solution (pH adjusted to 6.4; Wako Pure Chemical Industries, Japan).

注意:以上内容是从某篇研究文章中自动提取的,可能无法正确显示。



Q&A
请登录并在线提交您的问题
您的问题将发布在Bio-101网站上。我们会将您的问题发送给本研究方案的作者和具有相关研究经验的Bio-protocol成员。我们将通过您的Bio-protocol帐户绑定邮箱进行消息通知。