細胞組織消化常用的幾種酶的選擇

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直接從生物體獲取的組織，一般需要將其消化成單個細胞才能進行體外培養(yǎng)。這種直接從離體組織獲得的細胞，更接近于生物體內(nèi)的生活狀態(tài)，且生物性狀尚未發(fā)生很大改變，因此在藥物篩選、細胞移植、類器官培養(yǎng)、腫瘤研究等眾多領(lǐng)域備受歡迎。但組織消化過程中常遇到多種問題，例如消化不完全、細胞死亡率高等。如何克服這些問題，其實消化酶的選擇是關(guān)鍵。
本文將為您介紹一些常用的消化酶及其適用組織，并為您羅列多種正常和腫瘤組織消化方案，供您參考。

來了解組織消化過程中常用的酶吧～

1.胰蛋白酶
胰蛋白酶(Trypsin)是目前應(yīng)用最廣泛的消化試劑，通過作用于與賴氨酸或精氨酸相連接的肽腱，除去細胞間粘蛋白及糖蛋白，影響細胞骨架，從而使細胞分離。胰蛋白酶適用于消化細胞間質(zhì)較少的軟組織，如胚胎、上皮、肝、腎等組織。但對于纖維組織和較硬的癌組織的效果差，常常將胰蛋白酶與其他消化酶聯(lián)合使用。EDTA可以用來增強胰蛋白酶的水解功能，而血清則會影響胰蛋白酶的活性。

2.膠原酶
膠原酶(Collagenase)通過水解細胞間質(zhì)的脯氨酸，從而使細胞離散。膠原酶對膠原的消化作用很強，它僅對細胞間質(zhì)有消化作用而對細胞損害不大，因此適于消化分離纖維性組織和較硬的癌組織。鈣鎂離子和血清不會對膠原酶的活性及消化作用產(chǎn)生影響。
膠原酶可細分為Ⅰ、Ⅱ、Ⅲ、Ⅳ、Ⅴ型膠原酶，不同類型膠原酶適用范圍不同(詳見下表)。

膠原酶類型及適用范圍表

3.透明質(zhì)酸酶
透明質(zhì)酸酶(Hyaluronidase)通過隨機降解透明質(zhì)酸的β-N-乙酰己糖胺-[1-4]糖苷鍵，從而降低透明質(zhì)酸的活性，常與膠原酶等聯(lián)合使用，用于結(jié)締組織的分離。

4.脫氧核糖核酸酶Ⅰ
脫氧核糖核酸酶Ⅰ(DeoxyribonucleaseⅠ, DNaseⅠ)作用于細胞分離降解出的DNA，防止DNA導(dǎo)致的細胞凝集，對細胞完整性無破壞作用。通常配合膠原酶或透明質(zhì)酸酶等聯(lián)合使用，不會單獨使用。

5.中性蛋白酶
中性蛋白酶(Dispase)具有溫和的蛋白酶水解活性，不會損壞細胞膜的完整性，常與膠原酶等聯(lián)合使用，其分離纖維樣組織的效率高于上皮組織。

6.彈性蛋白酶
彈性蛋白酶(Elastase)通過作用于彈性蛋白的肽鍵、酰胺鍵和酯鍵將其水解，常與胰蛋白酶和膠原酶等聯(lián)合使用，用于分離結(jié)締組織和含有大量細胞網(wǎng)狀纖維的組織。

還有消化方案供您參考～
不同組織消化方案有所不同，同種組織的消化方案也不盡相同，考慮到組織的特殊性及酶的多樣性，針對具體組織仍需試驗摸索以確定最佳消化方案。以下列表方法供大家參考。

表一：正常組織消化用酶列表

表二：腫瘤組織消化用酶列表

參考文獻

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細胞組織消化常用的幾種酶的選擇