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Eppendorf 细胞培养皿
小转瓶1000ml
胶原包被75cm²培养瓶,250ml,密封盖,
[VIP第1年] 指数:2
通过静安区市场监管局认证 
Thyroglobulin Antibo
3T3-L1分化培养基，3T
小肠组织cDNA，胎儿
Human TGF beta 3 - H
Ach Thawing Medium
CKM Antibody
Human CD62P / P-Sel| 货号： | AVP004-3 |
| 保修期： | 30天 |
| 现货状态： | 期货 |
| 供应商： | Reinnervate |
| 规格： | 3 inserts |
Product Overview
Alvetex® is a highly porous polystyrene scaffold designed for 3D cell culture.
Cells grown in Alvetex possess a natural tissue-like structure that enables them to function in a more physiologically relevant manner. Alvetex 3D cell culture enables cells maintain their in vivo morphology, behavior and responsiveness within an in vitro model system.
Alvetex is a unique cellular environment
Manufactured to the highest standards of consistency each disc is engineered to a thickness of just 200 microns with pore sizes of 36-40 microns. No cell is ever further than 100 microns from the nutrient source enabling easily exchange of nutrients, gases and waste products by passive diffusion across short distances.
Say goodbye to risks of foreign unknown materials such as proteins or cytokines of animal origin – unlike conventional materials used in cell culture Alvetex is made from polystyrene and is completely inert.
Alvetex has been designed for simple and routine use. It uses conventional cell culture plasticware. Therefore any cell biologist can get into Alvetex 3D cell culture.
- No specialist equipment is required
- No changes to media and optimized cell feeding protocols
- Compatible with the majority of down stream analytical techniques.
Maintain natural cell shape and morphology
In conventional 2D cell culture, cells come into contact with the flat surface of the culture vessel (e.g. Petri dish, flask or multi-well plate). In this unnatural environment cells become flattened against the substrate. In this abnormally thin structure there has been significant re-modelling of the internal cellular components. The entire cytoskeleton is re-modelled and organelles such as the nucleus are flattened.
Well Insert Format
The presentation of Alvetex Scaffold in well insert formats is versatile, enabling long term 3D culture as cells can receive nutrients from media above and below the membrane, sustaining optimal 3D cell growth.
Currently there are two well insert sizes available: AVP004-3 (22 mm diameter) and AVP005-3 (15 mm diameter). Both are supplied in blister packs with three individually sealed inserts containing Alvetex Scaffold. The 6- and 12-well inserts are designed to fit into most 6-well plates or Reinnervate’s custom-made ‘Well Insert Holder in Deep Petri Dish’ (AVP015). Snapping the extended wings off AVP005-3 will also enable it to fit into a 12-well plate. Note that plates and well insert holders are not supplied with the product and have to be sourced separately.
什么是Alvetex®支架?
Alvetex® 系列产品是特别针对三维细胞培养设计的高度孔隙化（成孔率>90%），由惰性聚苯乙烯制成，厚度为200µm，孔径约为36-40µm的圆盘状（Disc）支架。
Fig 1. 扫描电镜（SEM）下观察到的Alvetex®支架微观结构
Fig 2. Alvetex®三维细胞培养支架系列产品类型一览：Well insert (6-well, 12-well)；Well plate (12-well, 24-well, 96-well, 384-well)；Well insert + Well insert holder in deep Petri-dish；Well insert + Perfusion plate.
为什么选择Alvetex®三维细胞培养支架?
Alvetex®支架为细胞生长提供了一个模拟体内生理条件的绝佳模型，细胞一旦接种到Alvetex®支架，即很容易进入到支架孔隙内并开始生长。所培养细胞在形态、细胞行为、反应特性方面都更接近于体内天然生长细胞。
Figure 3. 传统2D培养体系与Alvetex®三维细胞培养支架培养的成纤维细胞结构（细胞骨架、细胞核）对比。与传统的二维培养相比，三维条件下培养的细胞，细胞形态更生动立体，更接近体内自然状态。
Figure 4. Alvetex®系列三维细胞培养支架培养的4种不同来源的肝细胞。培养细胞经Bouin’s溶液固定、石蜡包埋后切片（10 µm），经两种染料——苏木精和伊红复染后置于光学显微镜下观察。（A）Alvetex® 6-well insert（AVP004）培养的HepG2细胞系（ATCC, HB-8065），40x物镜；（B）Alvetex® 24-well plate（AVP006）培养的Upcyte®肝细胞（Medicyte GmbH）， 20x物镜；（C）Alvetex® 12-well plate（AVP006）培养的冻存大鼠肝细胞（Biopredic Int.），40x物镜；（D） Alvetex® 12-well insert（AVP005）培养的大鼠肝细胞（Abcellute Ltd.）， 40x物镜；
Figure 5. 与传统2D培养体系相比，Alvetex®三维培养支架培养的HepG2细胞系表现出极佳的生长状态：细胞均一性提升，细胞间相互连接紧密度增加，并且开始形成较为复杂的细胞结构如微绒毛（microvilli）等。（A）HepG2细胞系在传统2D培养体系下培养7天的扫描电镜图片；（B）扫描电镜下观察到的于Alvetex® 三维培养支架内培养7天的HepG2细胞系；（C）高倍投射电镜下观察到的使用Alvetex®三维培养支架培养21天的HepG2细胞系，可以观察到微绒毛（microvill）、紧密连接（tight junctions）以及胆小管样结构（bile canaliculi-like structures）。
Figure 6. （左图）使用Alvetex®三维培养支架培养的HepG2细胞系，其细胞活性（MTT法检测）及分泌进入培养基的白蛋白（Albumin）水平显著高于传统2D模式下培养的HepG2细胞。（右图）以不同的接种密度（8x, 32x, and 64x 104 cells per well）将大鼠原代肝细胞接种于Alvetex®三维培养支架内培养24小时，使用钙黄绿素乙酰基甲基酯（calcein AM）/溴化乙锭（ethidium）异源二聚体标记法测定细胞活力，生长于Alvetex®三维培养支架内的原代肝细胞活性亦显著高于2D模式。
Figure 7. 与传统2D培养方式相比，Alvetex®三维培养支架培养的大鼠原代肝细胞在培养48h、72h后诱导产生不同亚型细胞色素氧化酶P450（CYP， CYP-1A2，CYP-2B1，CYP-3A2）的能力显著提升。
Alvetex®3D细胞培养支架操作流程
一、预处理
首先将Alvetex®支架完全浸没于70%酒精内润洗1次，吸弃酒精，PBS润洗支架2次。为了防止支架风干，请将支架浸没于第二次润洗时使用的PBS内，直至需要。
二、包被
为促进细胞粘着，选择合适的coating solution （poly-D or L-lysine，Fibronectin，Collagen I等）包被Alvetex®支架。以poly-D or L-lysine包被为例：吸弃PBS，加入poly-D or L-lysine，500/Well，盖好盖子，室温孵育1小时；
三、接种
将待培养细胞重悬于少量培养基内制成细胞悬液，吸弃培养板内多余的coating solution，然后小心地将细胞悬液转移接种至Alvetex®支架内，为保证最佳培养效果，请将细胞接种至支架最中央的位置。不同规格Alvetex®支架推荐接种密度及接种体积详见表1。
四、孵育
接种完毕后，将培养板的盖子盖上，放置于37℃，5% CO2培养箱内孵育至少30分钟以促进细胞附着到支架上。不同规格Alvetex®支架推荐孵育时间详见表1。
表1. 不同规格Alvetex®支架推荐接种密度、接种体积以及孵育时间
五、培养
孵育结束后，根据所培养的细胞类型、代谢活力等需求不同，轻轻地往培养板的每一个Well里面加入适量的培养基，然后再次置于37℃，5% CO2培养箱内孵育一段时间后，首次换液，之后每隔2-3天换液（据所培养的细胞类型、代谢活力等需求不同，首次换液时间以及换液频率有所不同）。不同规格Alvetex®支架推荐添加的培养基用量详见表2。
六、下游分析
传统2D培养模式下用于分析、检测的标准细胞及分子生物学方法如显微成像、免疫组织化学、免疫细胞化学、免疫荧光、细胞活力检测、蛋白质组学/基因组学定量分析等，同样适用于Alvetex®3D细胞培养支架培养的细胞。
Figure 8. A. CHO-K1细胞分别以0 cells, 0.5 million cells和2 million cells/支架的接种密度接种于Alvetex®支架（AVP002），培养24h后，使用无毒性的0.5 % 中性红染料（Sigma, N6264-50ML）染色并置于显微镜（明场）下观察；B. 人角质化细胞系（HaCaT）于Alvetex®支架培养7天，固定、石蜡包埋、切片后通过免疫荧光技术进行分析：phase contrast (top), blue fluorescent Hoescht 33258 nuclei stain (middle)，Ki67 staining (bottom).
Figure 9. GFP转染的CHO-K1细胞于Alvetex® 支架（AVP004）培养6天，激光共聚焦显微镜下观察到的单一的GFP转染CHO-K1细胞为期3个小时的细胞行为变化：获取图像间隔30min。
Figure 10. HepG2细胞系于Alvetex® 支架（AVP002）培养3天后，分别进行Hoechst 33342 (blue), cytokeratin 8 (green) 和Nile Red (red)标记并在激光共聚焦显微镜下观察。
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