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J Extracell Vesicles：使用荧光亲脂膜染料的囊泡吸收研究中的混杂因素

2017-12-03 MedSci MedSci原创

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小细胞外囊泡（sEV），如外泌体，是蛋白质、RNA和DNA的纳米载体。纯sEV的分离仍然具有挑战性，既往研究报道分离sEV的过程中常会受蛋白质和脂蛋白的污染。细胞摄取是了解exosome和sEV功能的基石，常使用亲脂性染料如PKH67或CellMask来标记囊泡。在这项研究中，我们探究了污染物是否可以混淆sEV和外泌体摄取实验的结果。尺寸排阻色谱法（SEC）从禁食或未禁食大鼠的血浆以及血清补充或无

小细胞外囊泡（sEV），如外泌体，是蛋白质、RNA和DNA的纳米载体。纯sEV的分离仍然具有挑战性，既往研究报道分离sEV的过程中常会受蛋白质和脂蛋白的污染。细胞摄取是了解exosome和sEV功能的基石，常使用亲脂性染料如PKH67或CellMask来标记囊泡。

在这项研究中，我们探究了污染物是否可以混淆sEV和外泌体摄取实验的结果。尺寸排阻色谱法（SEC）从禁食或未禁食大鼠的血浆以及血清补充或无血清条件细胞培养基中分离sEVs。使用纳米颗粒追踪，蛋白质和甘油三酯测定和免疫测定来表征洗脱液组分。使用不同的亲脂性染料标记SEC片段，使用内皮细胞或原代心肌细胞定量细胞摄取。

细胞染料转移与SEC中sEV的含量不相符，但是受脂蛋白和蛋白质含量的严重影响。大鼠的隔夜禁食降低了脂蛋白含量，并且也减少了染料转移，而晚期，SEV不足/富含蛋白质的部分表现出更大的染料转移。通过使用sEV消耗的血清或纯蛋白质样品的染色的实验清楚地显示在完全不存在sEV的情况下发生染料转移的可能性。

总之，蛋白质和脂蛋白可以对亲脂性染料向受体细胞的转移有所影响。考虑到sEV和外泌体分离物污染的可能性，应谨慎控制亲脂性染料染色实验，并谨慎解释结论。

原始出处：

Takov K, Yellon DM, Davidson SM. Confounding factors in vesicle uptake studies using fluorescent lipophilic membrane dyes. J Extracell Vesicles. 2017 Oct 12;6(1):1388731. doi: 10.1080/20013078.2017.1388731. eCollection 2017.

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    2018-07-12 shanyongle

    #ACE#

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    2018-05-02 zhaozhouchifen

    #Cell#

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    2018-09-25 bsmagic9140

    #TRA#

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    2018-09-17 ms6897892027066920

    #荧光#

    0

  5. [GetPortalCommentsPageByObjectIdResponse(id=1655814, encodeId=5833165581442, content=<a href='/topic/show?id=2a011829d7' target=_blank style='color:#2F92EE;'>#ACE#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=0, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=1829, encryptionId=2a011829d7, topicName=ACE)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=635024767990, createdName=shanyongle, createdTime=Thu Jul 12 16:27:00 CST 2018, time=2018-07-12, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1870307, encodeId=77cb18e0307b3, content=<a href='/topic/show?id=fd764459a8' target=_blank style='color:#2F92EE;'>#Cell#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=26, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=4459, encryptionId=fd764459a8, topicName=Cell)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=524d95, createdName=zhaozhouchifen, createdTime=Wed May 02 14:27:00 CST 2018, time=2018-05-02, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1656919, encodeId=1cf5165691992, content=<a href='/topic/show?id=3ca31e71903' target=_blank style='color:#2F92EE;'>#TRA#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=18, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=17719, encryptionId=3ca31e71903, topicName=TRA)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=757b24868428, createdName=bsmagic9140, createdTime=Tue Sep 25 06:27:00 CST 2018, time=2018-09-25, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1707775, encodeId=6bde1e07775ac, content=<a href='/topic/show?id=bd958e237a6' target=_blank style='color:#2F92EE;'>#荧光#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=21, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=87237, encryptionId=bd958e237a6, topicName=荧光)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=ae2931492970, createdName=ms6897892027066920, createdTime=Mon Sep 17 12:27:00 CST 2018, time=2018-09-17, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1960572, encodeId=3e3119605e278, content=<a href='/topic/show?id=dbe6445416' target=_blank style='color:#2F92EE;'>#CEL#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=27, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=4454, encryptionId=dbe6445416, topicName=CEL)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=c3ff68, createdName=维他命, createdTime=Sun Nov 11 07:27:00 CST 2018, time=2018-11-11, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1588289, encodeId=da021588289b8, content=<a href='/topic/show?id=1cb8e014e0' target=_blank style='color:#2F92EE;'>#ESI#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=25, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=7014, encryptionId=1cb8e014e0, topicName=ESI)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=f4f817593223, createdName=skhzy, createdTime=Tue Dec 05 05:27:00 CST 2017, time=2017-12-05, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1599824, encodeId=a2e71599824e3, content=<a href='/topic/show?id=be324039e31' target=_blank style='color:#2F92EE;'>#囊泡#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=17, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=40397, encryptionId=be324039e31, topicName=囊泡)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=86bc18759177, createdName=charl1234573, createdTime=Tue Dec 05 05:27:00 CST 2017, time=2017-12-05, status=1, ipAttribution=)]
    2018-11-11 维他命

    #CEL#

    0

  6. [GetPortalCommentsPageByObjectIdResponse(id=1655814, encodeId=5833165581442, content=<a href='/topic/show?id=2a011829d7' target=_blank style='color:#2F92EE;'>#ACE#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=0, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=1829, encryptionId=2a011829d7, topicName=ACE)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=635024767990, createdName=shanyongle, createdTime=Thu Jul 12 16:27:00 CST 2018, time=2018-07-12, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1870307, encodeId=77cb18e0307b3, content=<a href='/topic/show?id=fd764459a8' target=_blank style='color:#2F92EE;'>#Cell#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=26, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=4459, encryptionId=fd764459a8, topicName=Cell)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=524d95, createdName=zhaozhouchifen, createdTime=Wed May 02 14:27:00 CST 2018, time=2018-05-02, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1656919, encodeId=1cf5165691992, content=<a href='/topic/show?id=3ca31e71903' target=_blank style='color:#2F92EE;'>#TRA#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=18, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=17719, encryptionId=3ca31e71903, topicName=TRA)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=757b24868428, createdName=bsmagic9140, createdTime=Tue Sep 25 06:27:00 CST 2018, time=2018-09-25, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1707775, encodeId=6bde1e07775ac, content=<a href='/topic/show?id=bd958e237a6' target=_blank style='color:#2F92EE;'>#荧光#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=21, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=87237, encryptionId=bd958e237a6, topicName=荧光)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=ae2931492970, createdName=ms6897892027066920, createdTime=Mon Sep 17 12:27:00 CST 2018, time=2018-09-17, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1960572, encodeId=3e3119605e278, content=<a href='/topic/show?id=dbe6445416' target=_blank style='color:#2F92EE;'>#CEL#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=27, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=4454, encryptionId=dbe6445416, topicName=CEL)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=c3ff68, createdName=维他命, createdTime=Sun Nov 11 07:27:00 CST 2018, time=2018-11-11, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1588289, encodeId=da021588289b8, content=<a href='/topic/show?id=1cb8e014e0' target=_blank style='color:#2F92EE;'>#ESI#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=25, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=7014, encryptionId=1cb8e014e0, topicName=ESI)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=f4f817593223, createdName=skhzy, createdTime=Tue Dec 05 05:27:00 CST 2017, time=2017-12-05, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1599824, encodeId=a2e71599824e3, content=<a href='/topic/show?id=be324039e31' target=_blank style='color:#2F92EE;'>#囊泡#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=17, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=40397, encryptionId=be324039e31, topicName=囊泡)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=86bc18759177, createdName=charl1234573, createdTime=Tue Dec 05 05:27:00 CST 2017, time=2017-12-05, status=1, ipAttribution=)]
    2017-12-05 skhzy

    #ESI#

    0

  7. [GetPortalCommentsPageByObjectIdResponse(id=1655814, encodeId=5833165581442, content=<a href='/topic/show?id=2a011829d7' target=_blank style='color:#2F92EE;'>#ACE#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=0, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=1829, encryptionId=2a011829d7, topicName=ACE)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=635024767990, createdName=shanyongle, createdTime=Thu Jul 12 16:27:00 CST 2018, time=2018-07-12, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1870307, encodeId=77cb18e0307b3, content=<a href='/topic/show?id=fd764459a8' target=_blank style='color:#2F92EE;'>#Cell#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=26, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=4459, encryptionId=fd764459a8, topicName=Cell)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=524d95, createdName=zhaozhouchifen, createdTime=Wed May 02 14:27:00 CST 2018, time=2018-05-02, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1656919, encodeId=1cf5165691992, content=<a href='/topic/show?id=3ca31e71903' target=_blank style='color:#2F92EE;'>#TRA#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=18, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=17719, encryptionId=3ca31e71903, topicName=TRA)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=757b24868428, createdName=bsmagic9140, createdTime=Tue Sep 25 06:27:00 CST 2018, time=2018-09-25, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1707775, encodeId=6bde1e07775ac, content=<a href='/topic/show?id=bd958e237a6' target=_blank style='color:#2F92EE;'>#荧光#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=21, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=87237, encryptionId=bd958e237a6, topicName=荧光)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=ae2931492970, createdName=ms6897892027066920, createdTime=Mon Sep 17 12:27:00 CST 2018, time=2018-09-17, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1960572, encodeId=3e3119605e278, content=<a href='/topic/show?id=dbe6445416' target=_blank style='color:#2F92EE;'>#CEL#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=27, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=4454, encryptionId=dbe6445416, topicName=CEL)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=c3ff68, createdName=维他命, createdTime=Sun Nov 11 07:27:00 CST 2018, time=2018-11-11, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1588289, encodeId=da021588289b8, content=<a href='/topic/show?id=1cb8e014e0' target=_blank style='color:#2F92EE;'>#ESI#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=25, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=7014, encryptionId=1cb8e014e0, topicName=ESI)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=f4f817593223, createdName=skhzy, createdTime=Tue Dec 05 05:27:00 CST 2017, time=2017-12-05, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1599824, encodeId=a2e71599824e3, content=<a href='/topic/show?id=be324039e31' target=_blank style='color:#2F92EE;'>#囊泡#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=17, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=40397, encryptionId=be324039e31, topicName=囊泡)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=86bc18759177, createdName=charl1234573, createdTime=Tue Dec 05 05:27:00 CST 2017, time=2017-12-05, status=1, ipAttribution=)]
    2017-12-05 charl1234573

    #囊泡#

    0