Real-time confocal imaging of trafficking of nestin-expressing multipotent stem cells in mouse whiskers in long-term 3-D histoculture
Real-time confocal imaging of trafficking of nestin-expressing multipotent stem cells in mouse whiskers in long-term 3-D histoculture
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DOI：
10.1007/s11626-012-9514-z
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发表时间：
2012-05-01
影响因子：
2.1
通讯作者：
Hoffman, Robert M.
中科院分区：
文献类型：
--
作者：
Duong, Jennifer;Mii, Sumiyuki;Hoffman, Robert M.
我们之前已经证明，表达巢蛋白的多能毛囊干细胞位于毛囊隆突上方，并且可以在体外分化为神经元和其他细胞类型。当将表达nestin的毛囊干细胞移植到切断的坐骨神经或损伤的脊髓时，它们促进了预先存在的轴突的恢复。我们以前也证明了胡须毛囊在毛乳头（DP）和隆突区（BA）中含有表达巢蛋白的干细胞，但它们的起源是在BA中。在本研究中，我们建立了从巢蛋白驱动绿色荧光蛋白（ND-GFP）的转基因小鼠中分离的胡须的长期GelfoamA（R）组织培养技术。共聚焦成像用于监测ND-GFP表达干细胞在BA和DP之间的真实的时间运输，以确定干细胞的命运。在2周的时间内观察到干细胞从BA向DP区域运输，并在GelfoamA（R）上广泛生长，形成神经样结构。这种长期组织培养ND-GFP小鼠胡须的新方法将能够广泛研究表达nestin的毛囊多能干细胞的行为。
We have previously demonstrated that nestin-expressing multipotent hair follicle stem cells are located above the hair follicle bulge and can differentiate into neurons and other cell types in vitro. The nestin-expressing hair follicle stem cells promoted the recovery of pre-existing axons when they were transplanted to the severed sciatic nerve or injured spinal cord. We have also previously demonstrated that the whisker hair follicle contains nestin-expressing stem cells in the dermal papilla (DP) as well as in the bulge area (BA), but that their origin is in the BA. In the present study, we established the technique of long-term GelfoamA (R) histoculture of whiskers isolated from transgenic mice in which nestin drives green fluorescent protein (ND-GFP). Confocal imaging was used to monitor ND-GFP-expressing stem cells trafficking in real time between the BA and DP to determine the fate of the stem cells. It was observed over a 2-week period that the stem cells trafficked from the BA toward the DP area and extensively grew out onto GelfoamA (R) forming nerve-like structures. This new method of long-term histoculture of whiskers from ND-GFP mice will enable the extensive study of the behavior of nestin-expressing multipotent stem cells of the hair follicle.