An improved method for the isolation and culture of rat epidermal stem cells.
An improved method for the isolation and culture of rat epidermal stem cells.
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一种改进的大鼠表皮干细胞分离培养方法。
DOI：
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发表时间：
2013-10
期刊：
影响因子：
--
通讯作者：
Yang RH, Xie JL, Shu B, Liu XS, Chen XD, Ruan SB,
中科院分区：
文献类型：
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作者：
Yang RH, Xie JL, Shu B, Liu XS, Chen XD, Ruan SB,
使用涉及表皮干细胞 (ESC) 的新型治疗策略来治疗烧伤和损伤需要更好地了解这些细胞的生物学特性，特别是它们的分离和在培养物中维持其独特特性。本研究的目的是描述一种从胎鼠皮肤中分离假定的 ESC 并将其长期培养的改进方法。通过含有0.5%中性蛋白酶的酶消化从胎鼠皮肤中获得单一ESC悬浮液。通过在 IV 型胶原平板上快速粘附来收获靶细胞，并在复合 DMEM 中培养。初次分离后，将细胞在无 K-血清培养基中连续培养。达到70-80%汇合后，用0.25%胰蛋白酶在37℃消化5-10分钟，并以1:2的比例传代。培养的ESC生长良好，细胞活力超过98%。四天后，检测到含有 100-200 个细胞的克隆，显示出鹅卵石样特征。快速贴壁细胞的角蛋白 15、19 和 P63 呈阳性。百分之八十三的细胞表达β1整合素。生长曲线显示快速贴壁细胞处于指数生长期。本文描述的方案提供了一种简化且有效的方法来分离和维持胎鼠皮肤表皮干细胞的长期培养。该方法对于从目前可用的各种转基因大鼠系中分离和研究 ESC 应该很有价值。
The management of burns and injuries using novel treatment strategies involving epidermal stem cells (ESC) requires a better understanding of the biology of these cells, in particular, their isolation and the maintenance of their unique characteristics in culture. The purpose of this study was to describe an improved method for isolating putative ESC from fetal rat skin and to maintain them long term in culture. Single ESC suspensions were obtained from fetal rat skin by enzyme digestion containing 0.5% neutral protease. The target cells were harvested by rapid adherence on type IV collagen plates and were cultured in complex DMEM. After primary isolation, cells were continuously cultured in K-serum free medium. After reaching 70-80% confluence, the cells were digested with 0.25% trypsin at 37°C for 5-10 minutes, and passaged at a ratio of 1:2. The cultured ESC showed good growth, resulting in cell viability of over 98%. Four days later, clones containing 100-200 cells were detected, showing cobblestone-like characteristics. The rapidly adherent cells were positive for keratin 15, 19 and P63. Eighty three percent of cells expressed β1 integrin. The growth-curve showed that the rapidly adherent cells were in the exponential growth phase. The protocol described in this paper provides a simplified and effective method to isolate and maintain long-term culture of epidermal stem cells from fetal rat skin. This method should be valuable for isolating and studying ESC from various transgenic rat lines that are currently available.
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影响因子：
4
作者：
M. Michel;Natalie Török;M. Godbout;Marc Lussier;Pierrette Gaudreau;André Royal;Lucie Germain
通讯作者：
M. Michel;Natalie Török;M. Godbout;Marc Lussier;Pierrette Gaudreau;André Royal;Lucie Germain
影响因子：
3.7
作者：
Charlotte Triel;M. Vestergaard;L. Bolund;T. Jensen;U. Jensen
通讯作者：
Charlotte Triel;M. Vestergaard;L. Bolund;T. Jensen;U. Jensen
影响因子：
5.2
作者：
A. Terunuma;V. Kapoor;C. Yee;W. Telford;M. Udey;J. Vogel
通讯作者：
A. Terunuma;V. Kapoor;C. Yee;W. Telford;M. Udey;J. Vogel
影响因子：
15.9
作者：
Li, A;Pouliot, N;Kaur, P
通讯作者：
Kaur, P
DOI：
10.1111/j.1610-0387.2005.05071.x
发表时间：
2005-12
期刊：
JDDG: Journal der Deutschen Dermatologischen Gesellschaft
影响因子：
--
作者：
Karin Rzepka;Grit Schaarschmidt;Michael Nagler;J. Wohlrab
通讯作者：
Karin Rzepka;Grit Schaarschmidt;Michael Nagler;J. Wohlrab