馬薩諸塞州總醫(yī)院(MGH)的研究人員最近找到一種成功地體外培養(yǎng)肝細(xì)胞的方法，培養(yǎng)的肝細(xì)胞具有藥物毒性篩選功能,。
該研究發(fā)表在即將出版的PNAS雜志上,。研究報(bào)告詳細(xì)介紹了肝細(xì)胞如何在高氧條件和無動(dòng)物血清的條件下生長，并如何快速發(fā)揮正常肝臟所具有的功能,。
快速有效的篩選出具有毒副作用的藥物能顯著降低新藥問世的成本,。因?yàn)楦闻K對(duì)代謝過程和體內(nèi)藥物清除具有重要作用，因此,，篩選出肝臟毒性反應(yīng)的藥物是確保用藥安全的關(guān)鍵,。但動(dòng)物研究并不能完全預(yù)測(cè)肝臟毒性反應(yīng)，而在培養(yǎng)條件下的肝細(xì)胞又很快會(huì)失去代謝能力,。
之前有研究表明,，細(xì)胞培養(yǎng)中常用的動(dòng)物血清對(duì)培養(yǎng)的肝細(xì)胞具有干擾作用，而且體外培養(yǎng)的肝細(xì)胞所處的氧氣環(huán)境降低了10倍,。因此,，研究人員認(rèn)為高氧、無血清環(huán)境是確保培養(yǎng)肝細(xì)胞的關(guān)鍵,。
研究人員利用小鼠和人類的肝臟細(xì)胞進(jìn)行研究，發(fā)現(xiàn)在無血清和含95%氧氣的條件下,，肝細(xì)胞很快能恢復(fù)正常代謝能力,。這些培養(yǎng)的肝細(xì)胞還能或快或慢的進(jìn)行藥物清除，并且連續(xù)幾周維持高水平的代謝能力,。(生物谷Bioon.com)
生物谷推薦原始出處：
PNAS August 31, 2009, doi: 10.1073/pnas.0906820106
Oxygen-mediated enhancement of primary hepatocyte metabolism, functional polarization, gene expression, and drug clearance
Srivatsan Kidambia, Rubin S. Yarmusha, Eric Novikb, Piyun Chaob, Martin L. Yarmusha and Yaakov Nahmiasa,c,1
aCenter for Engineering in Medicine, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02111;
bHμREL Corporation, Beverly Hills, CA 90211; and
cThe Selim and Rachel Benin School of Computer Science and Engineering, The Hebrew University of Jerusalem, Jerusalem 91904, Israel
The liver is a major site for the metabolism of xenobiotic compounds due to its abundant level of phase I/II metabolic enzymes. With the cost of drug development escalating to over $400 million/drug there is an urgent need for the development of rigorous models of hepatic metabolism for preclinical screening of drug clearance and hepatotoxicity. Here, we present a microenvironment in which primary human and rat hepatocytes maintain a high level of metabolic competence without a long adaptation period. We demonstrate that co-cultures of hepatocytes and endothelial cells in serum-free media seeded under 95% oxygen maintain functional apical and basal polarity, high levels of cytochrome P450 activity, and gene expression profiles on par with freshly isolated hepatocytes. These oxygenated co-cultures demonstrate a remarkable ability to predict in vivo drug clearance rates of both rapid and slow clearing drugs with an R2 of 0.92. Moreover, as the metabolic function of oxygenated co-cultures stabilizes overnight, preclinical testing can be carried out days or even weeks before other culture methods, significantly reducing associated labor and cost. These results are readily extendable to other culture configurations including three-dimensional culture, bioreactor studies, as well as microfabricated co-cultures.
