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更新時間：2022-07-21 15:26:06瀏覽次數(shù)：229次
聯(lián)系我時，請告知來自 儀表網(wǎng)產(chǎn)品介紹：鑒于人肝細胞原代培養(yǎng)在藥物代謝、毒性、副作用的研究中被視為核心檢驗標(biāo)準(zhǔn)
鑒于人肝細胞原代培養(yǎng)在藥物代謝、毒性、副作用的研究中被視為核心檢驗標(biāo)準(zhǔn)，而原代培養(yǎng)的肝細胞經(jīng)常面臨諸如解毒基因表達量下調(diào)的問題（如膜轉(zhuǎn)運蛋白、結(jié)合酶、細胞色素P45），在Vinci B等人的肝細胞研究中（Vinci B, et al. Modular bioreactor for primary human hepatocyte culture: medium flow stimulates expression and activity of detoxification genes. Biotechnol J. 2011; 6: 554-64），認為正是傳統(tǒng)靜置培養(yǎng)無法模擬的某些生理刺激，引發(fā)了基因表達的下調(diào)。而Vinci B通過Quasi Vivo流動培養(yǎng)人原代肝臟細胞后，不論基因表達量、酶活還是生物參數(shù)，均證明流動培養(yǎng)可特異性地上調(diào)多種解毒酶基因的表達，而且流動培養(yǎng)的人貼壁肝細胞，其解毒酶基因家族的表達水平接近或高于新鮮分離的肝臟細胞。
實驗流程
1、將原代肝細胞進行靜態(tài)培養(yǎng)，在第七天分組，一組保持靜態(tài)培養(yǎng)，一組改為Quasi Vivo流動培養(yǎng)，繼續(xù)培養(yǎng)至二十一天，進行檢測。
2、分別進行流動培養(yǎng)、靜態(tài)培養(yǎng)、標(biāo)準(zhǔn)培養(yǎng)十四天后的肝臟細胞(FT297)，形態(tài)無差異
3、流動培養(yǎng)下的肝細胞CYP1A1、CYP1A2、CYP2B6、CYP3A4、GSTa等解毒酶基因表達更高，到達峰值時長有差異。
4、多種解毒基因上調(diào)，CYP1A1等基因表達甚至高于新鮮分離的肝臟細胞（FIH）
5、流動培養(yǎng)提高了肝臟細胞CYP3A4、UGT2B4/7、CYP2C9的活性，化合物代謝速率顯著提高。
文章小結(jié)：
1、人原代肝臟細胞分別進行7-21天的流動培養(yǎng)、靜態(tài)培養(yǎng)；
2、檢測肝臟細胞32個基因的表達量、酶活和生物參數(shù)；
3、使用流動培養(yǎng)可提高人貼壁肝臟細胞解毒基因表達：
①UGT（尿苷二磷酸葡萄糖醛酸轉(zhuǎn)移酶）基因家族表達上調(diào)：UGT1A1/UGT2B4/UGT2B7；
②多藥耐藥相關(guān)蛋白1 (MDR1)和MRP2基因的mRNA的表達上調(diào)；
③外源物質(zhì)/藥物代謝和運輸基因表達上調(diào)，并激活某些酶活性：CYP1A1/1A2/2B6/2C9/3A4；
④GST(S-轉(zhuǎn)移酶)基因表達上調(diào)。
用于(貼壁)細胞培養(yǎng)的Quasi Vivo流動培養(yǎng)系統(tǒng)，利用培養(yǎng)基循環(huán)流動，模擬血流剪切應(yīng)力環(huán)境，更貼近人肝臟細胞的體內(nèi)環(huán)境，能促進原代肝臟細胞增殖、貼壁肝細胞的基因表達等。作為英國Kirkstall公司核心產(chǎn)品，Quasi Vivo流動培養(yǎng)系統(tǒng)創(chuàng)新地模擬了體內(nèi)動態(tài)環(huán)境，并可同時結(jié)合3D培養(yǎng)構(gòu)建細胞模型。流動培養(yǎng)系統(tǒng)主要由高精度蠕動泵（用于培養(yǎng)基循環(huán)流動）、特殊設(shè)計的培養(yǎng)腔室（低通量培養(yǎng)用、中-高通量培養(yǎng)用）耗材組成。Quasi Viv設(shè)備可自行拼裝，操作方便，按產(chǎn)品說明書清洗和滅菌后，所有耗材可長期、重復(fù)使用，大大降低了Quasi Vivo的使用成本。
目前，Quasi Vivo流動培養(yǎng)系統(tǒng)已在超過70個專業(yè)研究機構(gòu)獲得應(yīng)用，已成功構(gòu)建包括呼吸系統(tǒng)（肺成纖維細胞、支氣管上皮細胞等）、肝臟、腎臟、心血管、腦組織、糖尿病等研究模型（原代細胞模型）。
Quasi Vivo參考文獻
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8.Ahluwalia A. et al., 2011. Hepatotoxicity of diclofenac in a Quasi-Vivo™ multicompartment bioreactor. oxicology Letters Volume: 205.
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29.Tommaso S. et al., 2011. Engineering Quasi-Vivo in vitro organ models. Advances in Experimental Medicine and Biology Volume: 745, pp 138-153.
30.Ahluwalia A. et al., 2011. Hepatotoxicity of diclofenac in a Quasi-Vivo™ multicompartment bioreactor. oxicology Letters Volume: 205.
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