中国组织工程研究与临床康复
中國組織工程研究與臨床康複
중국조직공정연구여림상강복
JOURNAL OF CLINICAL REHABILITATIVE TISSUE ENGINEERING RESEARCH
2010年
14期
2647-2651
,共5页
视网膜%视皮质%神经元%原代培养%大鼠
視網膜%視皮質%神經元%原代培養%大鼠
시망막%시피질%신경원%원대배양%대서
背景:神经元的原代培养是进行神经系统结构和功能研究的一种重要手段.如果能建立稳定良好的视网膜、视皮质神经细胞体外培养体系,对深入研究其各种细胞成分的生物学功能、病理改变过程与机制以及药物反应等十分重要.目的:对比观察原代培养新生大鼠视网膜及视皮质神经元的特点,探求最佳分离和培养方法.方法:分别采用机械分离及酶消化法分离新生大鼠视网膜及视皮质神经元,应用含体积分数为10%新牛牛血清、10%F-12 Nutrient Mixtures的DMEM培养基进行接种培养,含2%B-27 Serum-Free Supplements的Neurobasal Medium进行维持培养,利用尼氏染色进行神经元鉴定.结果与结论:培养的神经元生长良好,胞体饱满,突起长.尼氏染色示视网膜神经元比例大于90%,视皮质神经元比例大于50%.提示视网膜及视皮质神经元培养方法及生长特点存在不同,需采用不同的方法进行培养以获得高纯度的神经元.
揹景:神經元的原代培養是進行神經繫統結構和功能研究的一種重要手段.如果能建立穩定良好的視網膜、視皮質神經細胞體外培養體繫,對深入研究其各種細胞成分的生物學功能、病理改變過程與機製以及藥物反應等十分重要.目的:對比觀察原代培養新生大鼠視網膜及視皮質神經元的特點,探求最佳分離和培養方法.方法:分彆採用機械分離及酶消化法分離新生大鼠視網膜及視皮質神經元,應用含體積分數為10%新牛牛血清、10%F-12 Nutrient Mixtures的DMEM培養基進行接種培養,含2%B-27 Serum-Free Supplements的Neurobasal Medium進行維持培養,利用尼氏染色進行神經元鑒定.結果與結論:培養的神經元生長良好,胞體飽滿,突起長.尼氏染色示視網膜神經元比例大于90%,視皮質神經元比例大于50%.提示視網膜及視皮質神經元培養方法及生長特點存在不同,需採用不同的方法進行培養以穫得高純度的神經元.
배경:신경원적원대배양시진행신경계통결구화공능연구적일충중요수단.여과능건립은정량호적시망막、시피질신경세포체외배양체계,대심입연구기각충세포성분적생물학공능、병리개변과정여궤제이급약물반응등십분중요.목적:대비관찰원대배양신생대서시망막급시피질신경원적특점,탐구최가분리화배양방법.방법:분별채용궤계분리급매소화법분리신생대서시망막급시피질신경원,응용함체적분수위10%신우우혈청、10%F-12 Nutrient Mixtures적DMEM배양기진행접충배양,함2%B-27 Serum-Free Supplements적Neurobasal Medium진행유지배양,이용니씨염색진행신경원감정.결과여결론:배양적신경원생장량호,포체포만,돌기장.니씨염색시시망막신경원비례대우90%,시피질신경원비례대우50%.제시시망막급시피질신경원배양방법급생장특점존재불동,수채용불동적방법진행배양이획득고순도적신경원.
BACKGROUND:Primary culture of neurons is an important way to study the structure and functions of the nervous system.It is also important to explore pathomechanism and medicine reaction of some ophthalmology diseases.OBJECTIVE:To explore an optimal way to the separation and culture of retinal and visual cortical neurons in new-bom rats through comparative observation of different primary culture methods.METHODS:Retinal and visual cortical neurons isolated from new-born rats were firstly cultured in Dulbecco's modified Eagle's medium supplemented with 10% new-born calf serum and 10% F12 nutrient mixtures,followed by maintaining culture in Neurobasal medium containing 2% B27 serum-free supplements.Nissls staining was performed for neuron identification.RESULTS AND CONCLUSION:Cultured neurons grew well with plump cell bodies and long processes.Nissls staining showed that the purity of retinal neurons was greater than 90% and the proportion of visual cortical neurons was higher than 50%.The results suggested that there are some differences in culturing methods and growth characteristics of retinal and visual cortical neurons of new-born rats,accordingly,different culture methods are required to obtain high purity neurons.
