【Corning】354262 产品使用方法
Corning® Matrigel® Matrix High Concentration (HC), Phenol-Red Free, LDEV-free
（354262）
基底膜是体内细胞下方的薄层细胞外基质。Corning Matrigel Matrix High Concentration (HC) Phenol Red Free是从Engelbreth-Holm-Swarm（EHS）小鼠肉瘤中提取的可溶化基底膜制备物，该肿瘤富含细胞外基质蛋白。其主要成分是层粘连蛋白，其次是胶原蛋白IV、肝素硫酸酯蛋白多糖和接触素/巢蛋白。Corning Matrigel Matrix HC Phenol Red Free还含有TGF-β、表皮生长因子、胰岛素样生长因子、成纤维细胞生长因子、组织纤溶酶原激活剂等生长因子，这些生长因子在EHS肿瘤中自然存在。Corning Matrigel Matrix HC Phenol Red Free可有效促进正常和转化的依赖锚定的上皮细胞和其他细胞类型的附着和分化，包括神经元、肝细胞、Sertoli细胞、小鸡晶状体和血管内皮细胞。Corning Matrigel Matrix HC Phenol Red Free会影响成年大鼠肝细胞、血管内皮细胞以及小鼠和人乳腺上皮细胞的三维培养中基因表达。它是几种肿瘤细胞侵袭实验的基础，支持体内周围神经再生，并为体外和体内研究血管生成提供必要的基质。Corning Matrigel Matrix HC Phenol Red Free还支持免疫抑制小鼠体内人类肿瘤的繁殖。还可以用于移植未经分选的乳腺细胞，以及嵌入在Corning Matrigel Matrix中的分选的上皮亚群。这种基质也被用作癌症干细胞模型，并且研究表明可以增强肿瘤在体内的生长速度。
应用：
表 1 354262产品信息
溶解方法
与Corning Matrigel Matrix HC Phenol Red Free接触的所有培养器具或培养基都必须预先冷却/冰冷，因为Corning Matrigel Matrix HC Phenol Red Free在10°C以上会开始凝胶化。始终将Corning Matrigel Matrix保持在冰上。
将Corning Matrigel Matrix HC Phenol Red Free解冻的方法是将小瓶完全浸入冰中并放入4°C冰箱的后部过夜。一旦Corning Matrigel Matrix HC Phenol Red Free解冻，摇晃瓶子以确保物质均匀分散。始终将Corning Matrigel Matrix放在冰上。采用无菌技术处理。将解冻的Corning Matrigel Matrix HC Phenol Red Free小瓶放入无菌区域，喷洒70%乙醇于瓶盖上，并风干。
可以使用预冷的移液管轻轻吸取Corning Matrigel Matrix HC Phenol Red Free，以确保均匀性。将Corning Matrigel Matrix HC Phenol Red Free分装到管中，当Corning Matrigel Matrix HC Phenol Red Free堵塞移液枪头端并/或导致移液管测量不准确时，请更换吸头。
如果凝胶化的Corning Matrigel Matrix HC Phenol Red Free放置在4°C的冰中24-48小时，可以重新液化。
Corning Matrigel Matrix HC Phenol Red Free可以作为薄凝胶层（0.5 mm），在其上培养细胞。也可以在Corning Matrigel Matrix HC Phenol Red Free内培养细胞，使用1 mm的层厚。过度稀释会导致薄而非凝胶化的蛋白质层。这对于细胞附着可能是有用的，但在分化研究中可能不太有效。
注射方案
Corning Matrigel Matrix HC Phenol Red Free可以通过皮下注射到小鼠体内（Corning Matrigel Plug Assay）来评估不同化合物的体内血管生成活性。高蛋白浓度促进肿瘤的生长，并使Corning Matrigel Plug在注射后保持完整性。这使得注入的肿瘤和/或血管生成化合物局限于原位分析和/或未来切除。
注意：Corning Matrigel Matrix产品的蛋白质浓度是批次特异性的，并在分析证书上提供。为了获得一致的结果，通过计算所需的特定蛋白质浓度（mg/mL）来稀释Corning Matrigel Matrix产品。为了保持凝胶的一致性，建议不要将Corning Matrigel Matrix稀释至低于3 mg/mL。使用冰冷的无血清培养基来稀释Corning Matrigel Matrix。冰冷的培养基可以直接加入冷冻的Corning Matrigel Matrix HC Phenol Red Free小瓶中，并按照建议的方法进行解冻。通过上下移液或在冰中旋转小瓶混合。
在将Corning Matrigel Matrix HC Phenol Red Free和Corning Matrigel Matrix/细胞悬浮液注射到小鼠之前，保持它们尽可能冷却，但不要冷冻。在整个过程中，保持Corning Matrigel和Corning Matrigel Matrix/细胞悬浮液尽可能无菌非常重要。
● 对于每只接受注射的小鼠，在冰上将细胞（2x105个或更多）和Corning Matrigel Matrix HC Phenol Red Free混合在最终体积为0.5 mL的溶液中。
●细胞应尽可能以较小的体积存在。通常情况下，将250 µl冰冷的含有2x106cells/mL的培养基与250µl冰冷的Corning Matrigel Matrix HC Phenol Red Free混合。
●使用19G针（对于组织样本）或23G针（对于培养细胞）在裸鼠的皮下注射细胞。注射应迅速进行，以防止Corning Matrigel Matrix HC Phenol Red Free凝固。
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