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细胞组织消化常用的几种酶的选择

2021.3.02

直接从生物体获取的组织，一般需要将其消化成单个细胞才能进行体外培养。这种直接从离体组织获得的细胞，更接近于生物体内的生活状态，且生物性状尚未发生很大改变，因此在药物筛选、细胞移植、类器官培养、肿瘤研究等众多领域备受欢迎。但组织消化过程中常遇到多种问题，例如消化不完全、细胞死亡率高等。如何克服这些问题，其实消化酶的选择是关键。
本文将为您介绍一些常用的消化酶及其适用组织，并为您罗列多种正常和肿瘤组织消化方案，供您参考。

来了解组织消化过程中常用的酶吧～

1.胰蛋白酶
胰蛋白酶(Trypsin)是目前应用最广泛的消化试剂，通过作用于与赖氨酸或精氨酸相连接的肽腱，除去细胞间粘蛋白及糖蛋白，影响细胞骨架，从而使细胞分离。胰蛋白酶适用于消化细胞间质较少的软组织，如胚胎、上皮、肝、肾等组织。但对于纤维组织和较硬的癌组织的效果差，常常将胰蛋白酶与其他消化酶联合使用。EDTA可以用来增强胰蛋白酶的水解功能，而血清则会影响胰蛋白酶的活性。

2.胶原酶
胶原酶(Collagenase)通过水解细胞间质的脯氨酸，从而使细胞离散。胶原酶对胶原的消化作用很强，它仅对细胞间质有消化作用而对细胞损害不大，因此适于消化分离纤维性组织和较硬的癌组织。钙镁离子和血清不会对胶原酶的活性及消化作用产生影响。
胶原酶可细分为Ⅰ、Ⅱ、Ⅲ、Ⅳ、Ⅴ型胶原酶，不同类型胶原酶适用范围不同(详见下表)。

胶原酶类型及适用范围表

3.透明质酸酶
透明质酸酶(Hyaluronidase)通过随机降解透明质酸的β-N-乙酰己糖胺-[1-4]糖苷键，从而降低透明质酸的活性，常与胶原酶等联合使用，用于结缔组织的分离。

4.脱氧核糖核酸酶Ⅰ
脱氧核糖核酸酶Ⅰ(DeoxyribonucleaseⅠ, DNaseⅠ)作用于细胞分离降解出的DNA，防止DNA导致的细胞凝集，对细胞完整性无破坏作用。通常配合胶原酶或透明质酸酶等联合使用，不会单独使用。

5.中性蛋白酶
中性蛋白酶(Dispase)具有温和的蛋白酶水解活性，不会损坏细胞膜的完整性，常与胶原酶等联合使用，其分离纤维样组织的效率高于上皮组织。

6.弹性蛋白酶
弹性蛋白酶(Elastase)通过作用于弹性蛋白的肽键、酰胺键和酯键将其水解，常与胰蛋白酶和胶原酶等联合使用，用于分离结缔组织和含有大量细胞网状纤维的组织。

还有消化方案供您参考～
不同组织消化方案有所不同，同种组织的消化方案也不尽相同，考虑到组织的特殊性及酶的多样性，针对具体组织仍需试验摸索以确定最佳消化方案。以下列表方法供大家参考。

表一：正常组织消化用酶列表

表二：肿瘤组织消化用酶列表

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