Isolation and culture of endothelial cells, pericytes and perivascular resident macrophage-like melanocytes from the young mouse ear.
Isolation and culture of endothelial cells, pericytes and perivascular resident macrophage-like melanocytes from the young mouse ear.
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DOI：
10.1038/nprot.2013.033
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发表时间：
2013-04
期刊：
影响因子：
14.8
通讯作者：
中科院分区：
文献类型：
--
作者：
该方案描述了一种基于生长介质的方法，从P10到P15(P，出生后一天)的小鼠的血管纹中获得耳蜗内皮细胞(ECs)、周细胞(PC)和血管周围驻留的巨噬细胞样黑素细胞(PVM/ms)。该程序不涉及样品组织的机械或酶消化。血管纹外植体在生长介质中选择性培养，在7~10d内获得原代细胞系。该方法简单可靠，经两代传代后可获得纯度达90%的高质量内皮细胞、PVM/MS和PC。该方法适用于从体积小、解剖结构复杂的器官和组织(如内耳毛细血管)制备原代培养细胞。高纯度的原代细胞系使基于细胞培养的细胞-细胞相互作用、屏障控制功能和药物作用的体外建模成为可能。
This protocol describes a growth medium–based approach for obtaining cochlear endothelial cells (ECs), pericytes (PCs) and perivascular resident macrophage-like melanocytes (PVM/Ms) from the stria vascularis of mice aged between P10 and P15 (P, postnatal day). The procedure does not involve mechanical or enzymatic digestion of the sample tissue. Explants of stria vascularis, ‘mini-chips’, are selectively cultured in growth medium, and primary cell lines are obtained in 7–10 d. The method is simple and reliable, and it provides high-quality ECs, PVM/Ms and PCs with a purity >90% after two passages. This protocol is suitable for producing primary culture cells from organs and tissues of small volume and high anatomical complexity, such as the inner ear capillaries. The highly purified primary cell lines enable cell culture–based in vitro modeling of cell-cell interactions, barrier control function and drug action.
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影响因子：
1.4
作者：
JUHN, SK;RYBAK, LP
通讯作者：
RYBAK, LP
影响因子：
50.3
作者：
Seaman, Steven;Stevens, Janine;Croix, Brad St.
通讯作者：
Croix, Brad St.
DOI：
10.1073/pnas.76.10.5217
发表时间：
1979-01-01
影响因子：
11.1
作者：
FOLKMAN, J;HAUDENSCHILD, CC;ZETTER, BR
通讯作者：
ZETTER, BR
影响因子：
15.9
作者：
JAFFE, EA;NACHMAN, RL;MINICK, CR
通讯作者：
MINICK, CR
影响因子：
19.6
作者：
Rops, AL;van der Vlag, J;Berden, JH
通讯作者：
Berden, JH