細(xì)胞描述
Macrophages are cells differentiated from circulating bone marrow-derived monocytes. The main function of macrophages is to remove cellular debris and destroy invading pathogens. Mouse Hepatic Macrophages (MHMa), which are also known as Kupffer cells, reside within the lumen of liver sinusoids. MHMa protect the liver by responding to pathogens and metastatic cells, while tolerating harmless self and foreign antigens, which enter via blood flow through the portal vein and hepatic artery [1]. Recent studies have shown that hepatic macrophages play an important role in fibrosis, liver inflammation, fatty liver disease, and liver transplantation [2-4]. MHMa are an excellent model for studying macrophage functions under normal physiological and pathological conditions.
細(xì)胞凍存步驟
待細(xì)胞生長狀態(tài)良好時,可進(jìn)行細(xì)胞凍存。下面T25瓶為例; 1.細(xì)胞凍存時,棄去培養(yǎng)基后,PBS清洗瓶底1-2次后加入1ml胰酶,細(xì)胞變圓脫落后,加入2ml完全培養(yǎng)基終止消化,可使用血球計數(shù)板計數(shù)。 2.1000RPM離心5分鐘去掉上清。用血清重懸浮,加DMSO至最終濃度為10%。加入DMSO后迅速混勻,按每1ml的數(shù)量分配到凍存管中,注意凍存管做好標(biāo)識。本公司按每個凍存管細(xì)胞數(shù)目大于1X106個細(xì)胞凍存。3.將凍存管置于程序降溫盒中,放入-80度冰箱,至少2個小時以后轉(zhuǎn)入液氮灌儲存。記錄凍存管位置以便下次拿取。