外泌体之家 | 细胞外膜泡领域核心平台—exosomes & microvesicles—小膜泡大作用

标题: 干细胞培养上清液如何储存？ [打印本页]
作者: comfortable    时间: 2016-10-28 17:31
标题: 干细胞培养上清液如何储存？

我想用差速离心分离间充质干细胞的外泌体？需要200ml的培养上清，平常需要好几天才能收齐这么多量的上清，所以可能要放好几天，我该如何储存上清呢？文献（详情见后面）里说分离的可以在4摄氏度2天，-20摄氏度比较长时间。我想如果我把上清液冻在-20摄氏度会不会对外泌体有影响？那我应该如何来储存上清液呢？请大家谈谈意见或者自己的经验，谢谢大家。

3.2.2. Storage Conditions
Exosome sizes decrease by about 60% in storage at 37 °C for 2 days. The size of exosomes does not
change during the first two days of storage at 4 °C. However, a decrease in size is observed after  
3–4 days. When exosomes are stored at −20 °C, their size remains constant over a long period. Notably,
multiple cycles of deep-freezing and thawing do not affect the size of exosomes. Therefore, −20 °C is a
suitable temperature to store exosomes [27].

作者: hereissyk    时间: 2016-10-29 10:31
New insights regarding exosome preservation are
emerging. Previous reports have indicated that exosomal
size decreases by approximately 60% after storage at
37°C for 2 days. However, the original size is preserved
when exosomes are stored at -80°C for 2 days. It is also
possible to preserve exosomes in either culture medium
or phosphate-buffered saline at -80°C for longer periods
of time [32, 33]. However, exosomes isolated using the
ExoQuick kit are only stable for up to 18 h, even if they are
stored at -80°C [3]. Exosomes extracted by ultrafiltration
or UC will begin to degrade and release their contents
after 2 h of storage at 37°C [34]. Because the proteins and
nucleic acids in exosomes are relatively unstable, storage
at -80°C is recommended.
Shao, Y., et al. (2016). "The functions and clinical applications of tumor-derived exosomes." Oncotarget.
作者: fang    时间: 2016-10-31 09:26
     看师姐是冻到-20度保存。
作者: comfortable    时间: 2016-10-31 20:11
Johnny 发表于 2016-10-28 18:18
一次多样点细胞吧，冻存的效果不好

谢谢  你是冰冻过吗  -80的效果不好是吧
作者: comfortable    时间: 2016-10-31 20:11
hereissyk 发表于 2016-10-29 10:31
New insights regarding exosome preservation are
emerging. Previous reports have indicated that exoso ...

谢谢 看来要冻在-80摄氏度啊
作者: comfortable    时间: 2016-11-1 19:02
Johnny 发表于 2016-11-1 10:54
我试过，上清液冻存后会析出很多结晶

谢谢 那我放一天可以吗 比如今天的收集了 放4摄氏度 然后明天和明天一起收集的共同离心  这样是否可行
作者: comfortable    时间: 2016-11-1 19:21
fang 发表于 2016-10-31 09:26
看师姐是冻到-20度保存。

请问师姐提取冰冻是否会有结晶 这个怎么处理



欢迎光临 外泌体之家 | 细胞外膜泡领域核心平台—exosomes & microvesicles—小膜泡大作用 (http://exosome.com.cn/) Powered by Discuz! X3.3