亲爱的研友该休息了!由于当前在线用户较少,发布求助请尽量完整的填写文献信息,科研通机器人24小时在线,伴您度过漫漫科研夜!身体可是革命的本钱,早点休息,好梦!

Cell Dissociation from the Tongue Epithelium and Mesenchyme/Connective Tissue of Embryonic-Day 12.5 and 8-Week-Old Mice

间充质 上皮 胚胎干细胞 结缔组织 细胞生物学 间充质干细胞 细胞 生物 人口 细胞培养 入侵 病理 胚胎 原肠化 胚胎发生 生物化学 医学 遗传学 基因 环境卫生
作者
Wenxin Yu,Mohamed Ishan,Zhonghou Wang,Hong‐Xiang Liu
出处
期刊:Journal of Visualized Experiments [MyJoVE Corporation]
卷期号: (167)
标识
DOI:10.3791/62163
摘要

Cell dissociation has been an essential procedure for studies at the individual-cell level and/or at a cell-population level (e.g., single cell RNA sequencing and primary cell culture). Yielding viable, healthy cells in large quantities is critical, and the optimal conditions to do so are tissue dependent. Cell populations in the tongue epithelium and underlying mesenchyme/connective tissue are heterogeneous and tissue structures vary in different regions and at different developmental stages. We have tested protocols for isolating cells from the mouse tongue epithelium and mesenchyme/connective tissue in the early developmental [embryonic day 12.5 (E12.5)] and young adult (8-week) stages. A clean separation between the epithelium and underlying mesenchyme/connective tissue was easy to accomplish. However, to further process and isolate cells, yielding viable healthy cells in large quantities, and careful selection of enzymatic digestion buffer, incubation time, and centrifugation speed and time are critical. Incubation of separated epithelium or underlying mesenchyme/connective tissue in 0.25% Trypsin-EDTA for 30 min at 37 °C, followed by centrifugation at 200 x g for 8 min resulted in a high yield of cells at a high viability rate (>90%) regardless of the mouse stages and tongue regions. Moreover, we found that both dissociated epithelial and mesenchymal/connective tissue cells from embryonic and adult tongues could survive in the cell culture-based medium for at least 3 h without a significant decrease of cell viability. The protocols will be useful for studies that require the preparation of isolated cells from mouse tongues at early developmental (E12.5) and young adult (8-week) stages requiring cell dissociation from different tissue compartments.
最长约 10秒,即可获得该文献文件

科研通智能强力驱动
Strongly Powered by AbleSci AI
更新
大幅提高文件上传限制,最高150M (2024-4-1)

科研通是完全免费的文献互助平台,具备全网最快的应助速度,最高的求助完成率。 对每一个文献求助,科研通都将尽心尽力,给求助人一个满意的交代。
实时播报
1秒前
隐形曼青应助西瓜撞地球采纳,获得10
7秒前
9秒前
12秒前
dilli完成签到 ,获得积分10
22秒前
希望天下0贩的0应助Jasmine采纳,获得10
24秒前
36秒前
李爱国应助阿智采纳,获得10
38秒前
Lucas应助科研通管家采纳,获得10
40秒前
莘莘发布了新的文献求助10
42秒前
bkagyin应助莘莘采纳,获得10
53秒前
1分钟前
pjh发布了新的文献求助10
1分钟前
1分钟前
Raunio完成签到,获得积分10
1分钟前
1分钟前
1分钟前
zqq完成签到,获得积分0
1分钟前
2分钟前
19900420发布了新的文献求助10
2分钟前
2分钟前
2分钟前
2分钟前
西瓜撞地球完成签到,获得积分20
2分钟前
2分钟前
充电宝应助19900420采纳,获得10
2分钟前
NexusExplorer应助llm采纳,获得10
2分钟前
2分钟前
2分钟前
2分钟前
pjh发布了新的文献求助30
2分钟前
快乐小菜瓜完成签到 ,获得积分10
2分钟前
3分钟前
身法马可波罗完成签到 ,获得积分10
3分钟前
bkagyin应助pjh采纳,获得10
3分钟前
llm发布了新的文献求助10
3分钟前
pjh完成签到,获得积分10
3分钟前
3分钟前
zs完成签到 ,获得积分10
3分钟前
3分钟前
高分求助中
歯科矯正学 第7版(或第5版) 1004
Semiconductor Process Reliability in Practice 1000
Smart but Scattered: The Revolutionary Executive Skills Approach to Helping Kids Reach Their Potential (第二版) 1000
Nickel superalloy market size, share, growth, trends, and forecast 2023-2030 600
GROUP-THEORY AND POLARIZATION ALGEBRA 500
Mesopotamian divination texts : conversing with the gods : sources from the first millennium BCE 500
Days of Transition. The Parsi Death Rituals(2011) 500
热门求助领域 (近24小时)
化学 医学 生物 材料科学 工程类 有机化学 生物化学 物理 内科学 纳米技术 计算机科学 化学工程 复合材料 基因 遗传学 催化作用 物理化学 免疫学 量子力学 细胞生物学
热门帖子
关注 科研通微信公众号,转发送积分 3234546
求助须知:如何正确求助?哪些是违规求助? 2880894
关于积分的说明 8217297
捐赠科研通 2548495
什么是DOI,文献DOI怎么找? 1377792
科研通“疑难数据库(出版商)”最低求助积分说明 647999
邀请新用户注册赠送积分活动 623347