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細胞組織消化常用的幾種酶的選擇

瀏覽次數:4595 發布日期:2020-5-25  來源:本站 僅供參考,謝絕轉載,否則責任自負

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。這種直接從離體組織獲得的細胞,更接近于生物體內的生活狀態,且生物性狀尚未發生很大改變,因此在藥物篩選、細胞移植、類器官培養、腫瘤研究等眾多領域備受歡迎。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.透明質酸酶 
透明質酸酶(Hyaluronidase)通過隨機降解透明質酸的β-N-乙酰己糖胺-[1-4]糖苷鍵,從而降低透明質酸的活性,常與膠原酶等聯合使用,用于結締組織的分離。

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酸酶等聯合使用,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合使用,用于分離結締組織和含有大量細胞網狀纖維的組織。

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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