Question Detail
For digesting mouse adipose-derived stem cells, is collagenase or a mixed enzyme better?
Reagent Selection
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All Answers4
B
Bio8967
2026-09-18
Liberase或同类混合酶的优势是批间稳定、对细胞表面抗原保留更好,适合下游流式、单细胞测序或需要重复性的实验;缺点是价格高、货期要询代理,一般比单胶原酶贵。经费有限就把它留给正式实验,国产或普通胶原酶做预实验筛选条件。
试
试剂老张
2026-09-16
胶原酶+分散酶通常比单用胶原酶更适合脂肪组织,分散酶帮助解离基底膜和ECM,胶状物会少一些。我一般用胶原酶I或II为基础,加分散酶和DNase I,37℃摇40 min左右,之后用100 µm和40 µm滤网过滤,离心去油。活率和得率要放在同一脂肪量下比,不然很难评价。
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Flask2292
2026-09-12
建议设置三组平行:单胶原酶+DNase I、胶原酶+分散酶+DNase I、Liberase+DNase I,固定脂肪量、小鼠年龄/性别、消化体积、摇速和终止条件。比较活率、贴壁率、成脂/成骨分化或表面标记,别只看一个时间点。价格和货期方面,进口混合酶小包装通常1-4周,具体要问代理商,不要按网价备货。
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抗体研究员
2026-08-01
小鼠腹股沟脂肪提干细胞,单用胶原酶1小时得率低、胶状物多,常见原因是DNA释放和脂肪未充分剪碎,不一定是酶种类问题。建议脂肪剪成乳糜状,加DNase I,消化30-45 min,中间轻柔摇晃,不要等1小时到点才看。胶原酶浓度过高或时间过长都会压活率。
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