A protocol for differentiation of human intestinal Caco-2 cells in asymmetric serum-containing medium
A protocol for differentiation of human intestinal Caco-2 cells in asymmetric serum-containing medium
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DOI：
10.1016/j.tiv.2012.01.008
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发表时间：
2012-12-01
影响因子：
3.2
通讯作者：
Sambuy, Yula
中科院分区：
文献类型：
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作者：
Ferruzza, Simonetta;Rossi, Carlotta;Sambuy, Yula
人肠Caco-2细胞系仍然代表了吸收性肠上皮细胞的最佳体外模型，尽管其来源于结肠腺癌。接种在滤器插入物上的Caco-2细胞在培养物中经历自发分化过程，该过程导致在2至3周后形成单层极化细胞，其通过紧密连接偶联并表达小肠肠上皮细胞的几种形态和功能特征。通常用于Caco-2细胞分化的培养基在顶（AP）和基底外侧（BL）隔室中均含有胎牛血清（FBS）补充物。FBS作为细胞培养基补充剂的使用在科学和伦理方面经常受到越来越多的质疑。我们已经表明，与在AP和BL培养基中使用10%FBS补充剂的标准条件（不对称方案）相比，仅向BL培养基中添加血清（不对称方案）似乎足以允许Caco-2细胞分化，如通过形态学、单层渗透性和碱性磷酸酶活性所监测的。虽然没有消除FBS的使用，但仅在BL培养基中添加FBS导致分化的更多生理条件和其使用的显著减少。(C)2012爱思唯尔有限公司保留所有权利。
The human intestinal Caco-2 cell line still represents the best available in vitro model of absorptive enterocytes, despite its origin from a colon adenocarcinoma. Caco-2 cells seeded on filter inserts undergo in culture a process of spontaneous differentiation that leads to the formation, after two to three weeks, of a monolayer of polarized cell, coupled by tight junctions and expressing several morphological and functional features of small intestinal enterocytes. The medium normally used for differentiation of Caco-2 cells contains a supplement of foetal bovine serum (FBS) in both the apical (AP) and basolateral (BL) compartments. The use of FBS as cell culture media supplement has been frequently and increasingly questioned on scientific and also on ethical grounds. We have shown that addition of serum only to the BL medium (asymmetric protocol) appears to be sufficient to allow differentiation of Caco-2 cells, as monitored by morphology, monolayer permeability and alkaline phosphatase activity, compared to standard conditions using 10% FBS supplement in both AP and BL media (asymmetric protocol). Although not eliminating the use of FBS, its addition only in the BL medium results in more physiological conditions for differentiation and in a significant reduction of its use. (C) 2012 Elsevier Ltd. All rights reserved.