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1、细胞的原代培养(Cell primary culture)实验 十一Evaluation only.Evaluation only. Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0.Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0. Copyright 2004-2011 Aspose Pty Ltd.Copyright 2004-2011 Aspose Pty Ltd.背景资料原代培养(primary culture):从生物体取材进行培养
2、,中途不分割培养物、不更换培养物的生长器皿。细胞的生长条件: 无菌 温度 pH 渗透压 氧气 营养及生长调节因子Evaluation only.Evaluation only. Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0.Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0. Copyright 2004-2011 Aspose Pty Ltd.Copyright 2004-2011 Aspose Pty Ltd.背景资料培养细胞的 生长类型
3、贴附型成纤维细胞型细胞上皮型细胞游走型细胞多形型细胞悬浮型 Evaluation only.Evaluation only. Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0.Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0. Copyright 2004-2011 Aspose Pty Ltd.Copyright 2004-2011 Aspose Pty Ltd.1、成纤维细胞型: 胞体呈梭型或不规则三角形,中央有卵圆形核,胞质突起,生长时呈
4、放射状。 除真正的成纤维细胞外,凡由中胚层间充质起源的组织,如心肌、平滑肌、成骨细胞、血管 内皮等常呈本型状态。 培养中细胞的形态与成纤维类似时皆可称之为成纤维细胞。背景资料Evaluation only.Evaluation only. Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0.Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0. Copyright 2004-2011 Aspose Pty Ltd.Copyright 2004-201
5、1 Aspose Pty Ltd.2、上皮型细胞: 细胞呈扁平不规则多角形,中央有圆形核,细胞彼此紧密相连成单层膜。生长时呈膜状移动, 处于膜边缘的细胞总与膜相连,很少单独行动。 起源于内、外胚层的细胞如皮肤表皮及其衍生物、消化管上皮、肝胰、肺泡上皮等皆成上皮型形态。背景资料Evaluation only.Evaluation only. Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0.Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0. Co
6、pyright 2004-2011 Aspose Pty Ltd.Copyright 2004-2011 Aspose Pty Ltd.3、游走细胞型:呈散在生长,一般不连成片,胞质常突起,呈活跃游走或变形运动,方向不规则。此型细胞不稳定,有时难以和其他细胞相区别。4、多型细胞型:有一些细胞,如神经细胞难以确定其规律和稳定的形态,统归于此类。背景资料Evaluation only.Evaluation only. Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0.Created with Aspose.Slides f
7、or .NET 3.5 Client Profile 5.2.0.0. Copyright 2004-2011 Aspose Pty Ltd.Copyright 2004-2011 Aspose Pty Ltd.悬浮型: 胞体圆形,不贴于支持物上,呈悬浮生长。这类细胞容易大量繁殖。 见于少数特殊的细胞,如某些类型的癌细胞及白血病细胞。 背景资料Evaluation only.Evaluation only. Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0.Created with Aspose.Slides for
8、.NET 3.5 Client Profile 5.2.0.0. Copyright 2004-2011 Aspose Pty Ltd.Copyright 2004-2011 Aspose Pty Ltd.原代细胞培养的一般方法: 植块培养法(explant culture) 分离(散)细胞培养法(dissociated cell culture)背景资料Evaluation only.Evaluation only. Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0.Created with Aspose.Slide
9、s for .NET 3.5 Client Profile 5.2.0.0. Copyright 2004-2011 Aspose Pty Ltd.Copyright 2004-2011 Aspose Pty Ltd.一、心肌细胞、表皮细胞的培养二、淋巴细胞的培养Evaluation only.Evaluation only. Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0.Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0. Copyrigh
10、t 2004-2011 Aspose Pty Ltd.Copyright 2004-2011 Aspose Pty Ltd.心肌细胞、表皮细胞的原代培养Evaluation only.Evaluation only. Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0.Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0. Copyright 2004-2011 Aspose Pty Ltd.Copyright 2004-2011 Aspose Pt
11、y Ltd.实验目的 了解细胞体外培养的原理、基本方法,掌握无菌操作方法及注意事项。 学习观察体外培养细胞的形态及生长状况。Evaluation only.Evaluation only. Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0.Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0. Copyright 2004-2011 Aspose Pty Ltd.Copyright 2004-2011 Aspose Pty Ltd.实验原理 来自机
12、体的细胞、组织、器官; 类似体内的体外环境; 人工培养条件下生存、生长、繁殖。Evaluation only.Evaluation only. Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0.Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0. Copyright 2004-2011 Aspose Pty Ltd.Copyright 2004-2011 Aspose Pty Ltd.主要仪器: 纯水仪;离心机;CO2培养箱;电热干燥箱;高压蒸汽
13、消毒锅;过滤器 及0.22 m滤膜; 超净台;眼科剪、镊;培养瓶;青霉素 瓶;平皿;吸管;移液管;血球计数板实验用品 Evaluation only.Evaluation only. Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0.Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0. Copyright 2004-2011 Aspose Pty Ltd.Copyright 2004-2011 Aspose Pty Ltd.实验用品 实验材料:7
14、12日龄的鸡胚Evaluation only.Evaluation only. Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0.Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0. Copyright 2004-2011 Aspose Pty Ltd.Copyright 2004-2011 Aspose Pty Ltd.实验用品 实验试剂:(1) M199 培养液(2) 小牛血清(3) 青、链霉素(4 ) Hanks液(5) 5NaHCO3(6
15、) 2碘酒(7) 75酒精Evaluation only.Evaluation only. Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0.Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0. Copyright 2004-2011 Aspose Pty Ltd.Copyright 2004-2011 Aspose Pty Ltd.实验步骤 一、培养前的准备工作 (1) 清洗与消毒(2) 配制溶液(3) 预热溶液(4) 紫外线消毒(5) 洗手和
16、着装(6) 进入无菌室 Evaluation only.Evaluation only. Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0.Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0. Copyright 2004-2011 Aspose Pty Ltd.Copyright 2004-2011 Aspose Pty Ltd.实验步骤 二、植块培养法培养鸡心肌细胞(1) 配培养液:含抗生素的M199基础培养液 9 ml小牛血清 1 ml过滤除菌。 Evaluation only.Evaluation only. Created with Aspose.Slides for .NET 3.5 Client Profile 5.2.0.0.Created w