從無致病性實驗小鼠組織中分離得到BALB/c小鼠心肌成纖維細胞。小鼠原代心臟成纖維細胞在涂有明膠基溶液的T25組織培養(yǎng)瓶中生長0.5小時，一般在AcceGen完全生長培養(yǎng)基中培養(yǎng)3-7天。然后文化被擴展。裝運前，細胞從瓶中分離出來，立即冷凍保存在小瓶中。每瓶至少含有1x10^6細胞每毫升，并冷凍運送。小鼠心臟原代成纖維細胞對細菌、酵母、真菌和支原體呈陰性。免疫熒光染色檢測細胞標記物的表達，抗體為anti-FSP1/S100A4。在AcceGen規(guī)定的細胞培養(yǎng)條件下，細胞可以1:2的分裂比擴增2-5代。不建議反復(fù)冷凍和解凍細胞。細胞培養(yǎng)的標準生化程序包括測定細胞間相互作用、RT-PCR、Western blotting、免疫沉淀、免疫熒光染色、流式細胞術(shù)或產(chǎn)生用于所需研究應(yīng)用的細胞衍生物。

BALB/c Mouse Primary Cardiac Fibroblasts from AcceGen are isolated from tissue of pathogen-free laboratory mice. Mouse Primary Cardiac Fibroblasts are grown in T25 tissue culture flasks pre-coated with gelatin-based solution for 0.5 hour and incubated in AcceGen Culture Complete Growth Medium generally for 3-7 days. Cultures are then expanded. Prior to shipping, cells are detached from flasks and immediately cryo-preserved in vials. Each vial contains at least 1x10^6 cells per ml and is delivered frozen. Mouse Primary Cardiac Fibroblasts are negative for bacteria, yeast, fungi, and mycoplasma. Cells are tested for expression of marker using the antibody of anti-FSP1/S100A4 by immunofluorescence staining. Cells can be expanded for 2-5 passages at a split ratio of 1:2 under the cell culture conditions specified by AcceGen. Repeated freezing and thawing of cells is not recommended. Standard biochemical procedures performed with cell cultures include the assay of cell to cell interaction, RT-PCR, Western blotting, immunoprecipitation, immunofluorescent staining, flow cytometry or generating cell derivatives for desired research applications.

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