论文部分内容阅读
本实验室自1984年起至今一直从事神经细胞体外培养的研究.Burrow已发表了在含血清培养液中培养出了鸡胚心脏组织的游离心肌细胞.本文旨在将新生大鼠的背根神经节或迷走神经结状神经节分别与自身的分离心肌细胞进行联合培养并首次成功地建立了感觉神经支配心肌细胞的体外培养模型.为研究神经细胞与心肌细胞之间的相互作用提供新的途径.1材料与方法1.1背根神经节或迷走神经结状神经节的培养,采用出生后24小时内的Sprague-Daluey大鼠,性别不限.在无菌条件下用显微解剖镜下取出背根神经节或迷走神经结状神经节,放入Tyrode平衡盐溶液中精细解剖,剥去神经节外膜,并以Tyrode平衡盐溶液反复冲洗,将其种植于预先涂有鼠尾胶的直径为24mm的圆玻片上同时可种3～4个背根神经节或迷走神经结状神经节,加培养液,然后将圆玻片放入小培养血中,置于37C,5%CO_2培养箱内约2～3小时,让组织块贴壁于涂有鼠尾胶的圆玻片上.1.2心肌细胞培养 无菌条件下,在取神经节的同时取出该动物的心脏,用D—Hank液将心脏漂洗干净.用眼科尖剪刀将其的成直径约1mm大小的小块,用0.125%胰酶消化5分钟,将该细胞悬液用210目的尼龙
This laboratory has been engaged in the research of neural cell culture in vitro since 1984. Burrow has published free cardiomyocytes cultured in serum-containing medium of chick embryo heart tissue.This paper aims to regenerate the dorsal root ganglion Ganglion or vagus nerve nodose ganglion separately from their own isolated cardiomyocytes and successfully established an in vitro culture model of sensory innervation cardiomyocytes for the first time.It provides a new way to study the interaction between nerve cells and cardiomyocytes. 1 Materials and Methods 1.1 dorsal root ganglion or vagus nodosum ganglion culture, using Sprague-Daluey rats within 24 hours after birth, gender is not limited to under aseptic conditions with a micro dissection removed the dorsal root nerve Section or vagus nerve nodose ganglion, placed in Tyrode balanced salt solution in fine dissection, stripped of the outer membrane of the ganglion, and repeated rinse with Tyrode balanced salt solution, it is planted in the mouse tail gel coated in advance with a diameter of 24mm round Slides on the same time can be planted 3 to 4 dorsal root ganglion or vagus nerve nodal ganglia, plus broth, and then round glass into small culture of blood, placed in 37C, 5% CO 2 incubator About 2 to 3 hours, so that the tissue block attached to the mouse tail coated round glass slide 1.2.2 cardiomyocyte culture aseptic conditions, taking the ganglion at the same time remove the heart of the animal with D-Hank solution to the heart Rinsed with ophthalmic sharp scissors to the size of about 1mm in diameter small pieces, with 0.125% trypsin digestion for 5 minutes, the cell suspension with 210 mesh nylon