目的利用牙囊细胞和成釉细胞贴壁速度及酶消化分离速度不同的特点,建立一种简便、快速纯化大鼠牙囊细胞的方法。方法取新生5~6 d SD大鼠下颌第一磨牙牙胚,在体视显微镜下剥离牙囊及成釉器,剪碎后酶消化并混合培养,再利用差速贴壁法和差...目的利用牙囊细胞和成釉细胞贴壁速度及酶消化分离速度不同的特点,建立一种简便、快速纯化大鼠牙囊细胞的方法。方法取新生5~6 d SD大鼠下颌第一磨牙牙胚,在体视显微镜下剥离牙囊及成釉器,剪碎后酶消化并混合培养,再利用差速贴壁法和差速传代法纯化牙囊细胞。结果原代细胞为牙囊细胞和成釉器细胞混合生长,差速传代培养到第2~3代可获得纯化的牙囊细胞。倒置显微镜下观察牙囊细胞呈梭形或三角形,免疫组织化学染色显示抗波形丝蛋白阳性,抗角蛋白阴性。结论双向差速法是一种高效、简便的纯化牙囊细胞的方法。展开更多
In this paper, the distribution and expression of diapause hormone (DH) at mRNA and protein levels in the central nervous system of Bombyx mori embryo and larva were studied using whole-mount in situ hybridization and...In this paper, the distribution and expression of diapause hormone (DH) at mRNA and protein levels in the central nervous system of Bombyx mori embryo and larva were studied using whole-mount in situ hybridization and immunocytochemistry. Whole-mount immunocytochemistry revealed that the distribution of the DH-like immunoreactivity was throughout the central nervous system including the brain, suboesophageal ganglion (SG) and thoracic ganglia(TG); and that the corpus cardiacum and terminal abdominal ganglion may be the site for DH release due to the presence of strong immunoreactivity. In situ hybridization with the probe labeled by digoxigenin shows that the BomDHmRNA was also localized in the mandibular, maxillary, labial cell clusters. In addition, a pair of lateral neurons in the SG and a pair of ventral midline neurons in each TG expressing the Bom-DH transcript were also identified. Thesere sults were consistent with the localization of Bom-DH mRNA in larva by in situ hybridization and the distribution of the gene by RT-PCR, which is some different from the results reported previously.展开更多
文摘目的利用牙囊细胞和成釉细胞贴壁速度及酶消化分离速度不同的特点,建立一种简便、快速纯化大鼠牙囊细胞的方法。方法取新生5~6 d SD大鼠下颌第一磨牙牙胚,在体视显微镜下剥离牙囊及成釉器,剪碎后酶消化并混合培养,再利用差速贴壁法和差速传代法纯化牙囊细胞。结果原代细胞为牙囊细胞和成釉器细胞混合生长,差速传代培养到第2~3代可获得纯化的牙囊细胞。倒置显微镜下观察牙囊细胞呈梭形或三角形,免疫组织化学染色显示抗波形丝蛋白阳性,抗角蛋白阴性。结论双向差速法是一种高效、简便的纯化牙囊细胞的方法。
文摘In this paper, the distribution and expression of diapause hormone (DH) at mRNA and protein levels in the central nervous system of Bombyx mori embryo and larva were studied using whole-mount in situ hybridization and immunocytochemistry. Whole-mount immunocytochemistry revealed that the distribution of the DH-like immunoreactivity was throughout the central nervous system including the brain, suboesophageal ganglion (SG) and thoracic ganglia(TG); and that the corpus cardiacum and terminal abdominal ganglion may be the site for DH release due to the presence of strong immunoreactivity. In situ hybridization with the probe labeled by digoxigenin shows that the BomDHmRNA was also localized in the mandibular, maxillary, labial cell clusters. In addition, a pair of lateral neurons in the SG and a pair of ventral midline neurons in each TG expressing the Bom-DH transcript were also identified. Thesere sults were consistent with the localization of Bom-DH mRNA in larva by in situ hybridization and the distribution of the gene by RT-PCR, which is some different from the results reported previously.
