The human squamous oesophagus has widespread capacity for clonal expansion from cells at diverse stages of differentiation.
The human squamous oesophagus has widespread capacity for clonal expansion from cells at diverse stages of differentiation.
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DOI：
10.1136/gutjnl-2013-306171
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发表时间：
2015-01
期刊：
影响因子：
24.5
通讯作者：
Fitzgerald RC
中科院分区：
文献类型：
--
作者：
Barbera M;di Pietro M;Walker E;Brierley C;MacRae S;Simons BD;Jones PH;Stingl J;Fitzgerald RC
在稳态和慢性损伤后，人类鳞状食管内环境稳定的细胞机制的知识是有限的。我们的目的是通过使用功能性3D方法更好地理解这些机制。通过免疫荧光法在3D上皮全标本上对10例正常鳞状食管中的增殖、有丝分裂和祖细胞谱系标志物的表达进行了评估。使用流式细胞术分选分离表达不同水平的上皮和祖细胞标志物的细胞，并通过qPCR和IF进行表征。通过集落形成细胞测定和体外器官型培养模型研究了它们的自我更新潜力。增殖和有丝分裂活性在乳头间基底层最高，并向乳头尖端线性下降（p<0.0001）。有丝分裂的方向是随机的整个基底层，不对称分裂并不局限于特定的细胞室。基于上皮和祖细胞标志物（CD34和EpCAM）的表达分选成不同群体的细胞在2D培养中的自我更新方面没有差异，无论是作为整个群体还是作为单细胞。在3D器官型培养物中，所有细胞亚型都能够重现起源组织的结构，并且决定3D培养物成功的主要因素是接种的细胞数量，而不是细胞类型。食管上皮细胞具有显著的自我更新可塑性。这种情况可以被视为离体创伤反应，并与最近在小鼠模型中的发现相一致。
Knowledge of the cellular mechanisms involved in homeostasis of human squamous oesophagus in the steady state and following chronic injury is limited. We aimed to better understand these mechanisms by using a functional 3D approach. Proliferation, mitosis and the expression of progenitor lineage markers were assessed in normal squamous oesophagus from 10 patients by immunofluorescence on 3D epithelial whole mounts. Cells expressing differential levels of epithelial and progenitor markers were isolated using flow cytometry sorting and characterised by qPCR and IF. Their self-renewing potential was investigated by colony forming cells assays and in vitro organotypic culture models. Proliferation and mitotic activity was highest in the interpapillary basal layer and decreased linearly towards the tip of the papilla (p<0.0001). The orientation of mitosis was random throughout the basal layer, and asymmetric divisions were not restricted to specific cell compartments. Cells sorted into distinct populations based on the expression of epithelial and progenitor cell markers (CD34 and EpCAM) showed no difference in self-renewal in 2D culture, either as whole populations or as single cells. In 3D organotypic cultures, all cell subtypes were able to recapitulate the architecture of the tissue of origin and the main factor determining the success of the 3D culture was the number of cells plated, rather than the cell type. Oesophageal epithelial cells demonstrate remarkable plasticity for self-renewal. This situation could be viewed as an ex vivo wounding response and is compatible with recent findings in murine models.
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影响因子：
50.3
作者：
Quante M;Bhagat G;Abrams JA;Marache F;Good P;Lee MD;Lee Y;Friedman R;Asfaha S;Dubeykovskaya Z;Mahmood U;Figueiredo JL;Kitajewski J;Shawber C;Lightdale CJ;Rustgi AK;Wang TC
通讯作者：
Wang TC
DOI：
10.1073/pnas.0909797107
发表时间：
2010-02-02
影响因子：
11.1
作者：
Saadi, Amel;Shannon, Nicholas B.;Fitzgerald, Rebecca C.
通讯作者：
Fitzgerald, Rebecca C.
影响因子：
29.4
作者：
Pan, Qiuwei;Nicholson, Anna M.;Jankowski, Janusz A.
通讯作者：
Jankowski, Janusz A.
影响因子：
4.6
作者：
Legg, J;Jensen, UB;Watt, FM
通讯作者：
Watt, FM
影响因子：
64.5
作者：
JONES, PH;WATT, FM
通讯作者：
WATT, FM