体外下丘脑神经细胞培养技术探讨.pdfVIP

体外下丘脑神经细胞培养技术探讨 陕西医学高等专科学校组织学与胚胎学教研室 (西安 710068) 王　兰　郑慧媛　宋天保3 　赵　璇　石明娟　王多宁3 3 　杨　颖3 3 　孟　洁 　　摘　要　目的: 探讨体外下丘脑神经细胞培养的技术。方法: 选用生后 1～ 3d 的 SD 大 鼠, 取下丘脑组织进行单细胞原代培养。结果: 培养 1d, 神经元胞体小, 圆形或梭形, 无突起 或突起极短; 培养 3d 时, 胞体明显增大, 突起长度增加; 到培养 7d, 神经元的胞体截面积和 突起长度均达到最高峰, 分别为 161. 63±21. 17Λm 2 和 103. 51±21. 33Λm; 到 14d 时, 神经 元的胞体截面积和突起长度基本维持在 7d 时的水平。结论: 培养前 7d 是下丘脑神经元体外 生长成熟的关键时期, 这种良好的发育状态可维持到 14d。体外下丘脑神经细胞培养技术的 关键在于准确的取材部位, 良好的单细胞悬液的制备, 尤其是贴壁物的选择。 　　主题词　细胞培养　神经元　下丘脑　体外研究 On techn iques for culture of nerve cells from hypotha lam us D epartm en t of H isto logy and Em b ryo logy, Shaanx iM edica l Co llege (X i’an 710068)　W ang L an　Zheng H u iyuan　Song T ianbao　et a l 　 　 ABSTRACT 　O bject ive: To find ou t the best techn iques fo r cu ltu re of nerve cells from hypo thalam us. M ethods: R ats aged 1～ 3 po stnatal days w ere used. T issue b lock s from hypo thalam usw ere removed and dispersed cells w ere cu ltu red in vitro. R esu lts: T he p rocesses of nerve cells iso lated from the rat hypo thalam us w ere found as early as on the first day of cu ltu re. D uring 1～ 3 days in cu ltu re neu rons grew fastest. T he cell body area and p rocess length of nerve cells reached their h ighest levels (161. 63± 21. 17Λm 2 and 103. 51±21. 33Λm , respect ively)on the 7th day. O n the 14th day, the som a size and p rocess length w ere m ain tained at the level of the 7th day. Conclu sion: T he first w eek is the crit ical period of in vitro developm en t and m atu rat ion of nerve cells from hypo thalam us, and their w ell2developed state m ain tain s th roughou t the second w eek. T he crit ical techn iques fo r cu ltu re of hypo thalam al nerve cells are accu rately gain ing the t issue, w ell p reparing the dispersed single2cell su spension, and especially select ing the p roper adhering substrate. 　　KEY WORD S　Cell　cu ltu re　N eurons　H ypo thalam us　 In vitro 　　下丘脑是神经内分泌中心, 它通过与垂体的 密切联系, 调节机体体温、摄食、生殖、水盐平衡等 内分泌活动。我们分别于 1999, 2002 年两次承担 省卫生厅科研课题中, 在关于一氧化氮 (N it ric ox ide, NO ) 和 一 氧 化 氮 合