西部戰(zhàn)區(qū)總醫(yī)院藥學部,在2021年發(fā)表文章到Cell Communication and Signaling的學術期刊中科研者使用美國 Invigentech轉染試劑,高效率轉染原代破骨細胞前體細胞(Osteoclast precursor cells),具體轉染原代破骨細胞前體細胞方法經驗如下:(注意:本文轉染實驗中使用的細胞數(shù)量、試劑用量和操作方法不適用于傳統(tǒng)lipo3000轉染試劑或類似產品)。
Cell culture
Sorted primary CD115(+) precursors were cultured in α-minimal essential medium (MEM) containing 10% FBS,1% penicillin-streptomycin solution and 50 ng ml? 1 colony stimulating factor (M-CSF, Abcam). After 3 days,the medium was replaced with fresh medium. LA was added at a concentration of 100 μg/ml to stimulate the precursors. To inhibit the expression of Cadherin-11, the primary osteoclast precursors were transfected with Cadherin-11 siRNA using INVI DNA RNA Transfection reagent (Invigentech)for 24h. The sequence of Cadherin-11 siRNA was CCAATGGACCAAGATTTAT. In some experiments, primary cells were treated with BYL719 (2.5 μM), GSK690693 (25 nM), 666–15 (50 nM), rapamycin (20 μM), or AMG-487 (30 μM).
美國 Invigentech介紹
美國 Invigentech(英克)公司是開發(fā)用于細胞培養(yǎng)的胎牛血清、細胞活力檢測、細胞篩選、細胞支原體清除、細胞轉染和組織再生的新型血清替代品全球之一;提供inviCELL™Platelet lysate無動物血清產品用于支持廣泛的細胞擴增和生產,包括培養(yǎng)間充質干細胞和多種免疫細胞系等,為制藥公司或者生物技術公司提供無血清細胞培養(yǎng)規(guī)模生產服務;INVI DNA RNA Transfection Reagent為原代細胞、懸浮細胞、小動物活體轉染提供高品質轉染試劑;此外我們引進的新設施可生產高純度注射用水和生物處理解決方案,用于滿足任何細胞、科研和企業(yè)客戶需求,保證每個批次胎牛血清、細胞活力檢測、細胞篩選、細胞轉染等產品質量的穩(wěn)定性,以加速安全有效的基于細胞和組織的治療研究、開發(fā)和商業(yè)化。
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