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10:03 min
January 30th, 2021
DOI :
10.3791/62182-v
Chapters
0:04
Introduction
0:27
Isolation and Culture of Human Keratinocytes from Neonatal Foreskin Tissue
2:37
Generating Skin Epidermosphere Cultures in Vitro
5:02
Epidermal Spheroid Re-Plating Assay
5:42
Characterization of Spheroid-Derived (SD) Sub-Populations by FACS
8:06
Results: Characterization of Epidermal Spheroid Formation and Spheroid-Derived NHKs
9:30
Conclusion
Transcript
本协议提出了培养和维持表皮球体培养的新方法,以及使用球体重新镀质检测密切研究这些培养和维持的策略。首先准备文本手稿中描述的洗涤介质。在层压流罩中,用五毫升的洗涤介质在五毫升圆锥管中两次清洗新生儿前皮,然后将洗过的前体转移到无菌的培养皿中。
使用手术刀和钳子,刮掉皮肤层的脂肪组织和松散的结缔组织,然后用洗涤介质重新洗净前皮。将包皮表皮侧放在一个六井板中,其中含有在洗涤介质中稀释的两毫升迪斯帕酶。将盘子转移到孵化器上四个小时。
孵化后,将前皮转移到培养皿中,并使用细尖钳
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Summary
在这里,我们描述了在3D悬架培养中系统地培养表皮球体的协议。本协议具有广泛的应用,用于各种上皮组织类型和几个人类疾病和条件的建模。
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建立高通量表皮球体培养系统,以模拟角膜细胞干细胞可塑性
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