Development of an enhanced human gastrointestinal epithelial culture system to facilitate patient-based assays.
Development of an enhanced human gastrointestinal epithelial culture system to facilitate patient-based assays.
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DOI：
10.1136/gutjnl-2013-306651
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发表时间：
2015-06
期刊：
影响因子：
24.5
通讯作者：
Stappenbeck TS
中科院分区：
文献类型：
--
作者：
VanDussen KL;Marinshaw JM;Shaikh N;Miyoshi H;Moon C;Tarr PI;Ciorba MA;Stappenbeck TS
人类肠上皮细胞的生长技术正在迅速发展。一个令人兴奋的可能性是，该系统可以作为个性化医疗和研究的平台。然而，为了实现这一目标，必须增强人上皮培养，使得来自个体的活组织检查可以用于在短时间范围内可重复地产生细胞系，使得可以进行多种功能性测定（即，屏障功能和宿主-微生物相互作用）。我们创建了一个大的面板的人胃肠道上皮细胞系（n = 65），从患者活检在常规的上，下内窥镜检查程序。使用含有生长关键因子（Wnt 3a、R-spondin和Noggin）的高浓度条件培养基快速扩增增殖性干/祖细胞。使用较低条件培养基浓度和Notch抑制的组合来分化这些细胞用于另外的测定。我们在培养的两周内从所有可接近的组织部位获得上皮细胞系。肠细胞系富含干细胞标志物，并快速生长为球状体，需要每3天以1：3-1：4传代。在分化条件下，肠上皮球状体表现出成熟上皮谱系的区域特异性发育。这些细胞在Transwell膜上生长时形成功能性极化单层，被分泌的粘液层覆盖。使用二维培养，这些细胞还表现出与各种致病性大肠杆菌菌株的新型粘附表型。该培养系统将促进人肠上皮细胞的个体间功能研究，包括宿主-微生物相互作用。
The technology for the growth of human intestinal epithelial cells is rapidly progressing. An exciting possibility is that this system could serve as a platform for individualized medicine and research. However, to achieve this goal, human epithelial culture must be enhanced so that biopsies from individuals can be used to reproducibly generate cell lines in a short time frame so that multiple, functional assays can be performed (i.e., barrier function and host-microbial interactions). We created a large panel of human gastrointestinal epithelial cell lines (n = 65) from patient biopsies taken during routine upper and lower endoscopy procedures. Proliferative stem/progenitor cells were rapidly expanded using a high concentration of conditioned media containing the factors critical for growth (Wnt3a, R-spondin and Noggin). A combination of lower conditioned media concentration and Notch inhibition was used to differentiate these cells for additional assays. We obtained epithelial lines from all accessible tissue sites within two weeks of culture. The intestinal cell lines were enriched for stem cell markers and rapidly grew as spheroids that required passage at 1:3–1:4 every 3 days. Under differentiation conditions, intestinal epithelial spheroids showed region-specific development of mature epithelial lineages. These cells formed functional, polarized monolayers covered by a secreted mucus layer when grown on Transwell membranes. Using two-dimensional culture, these cells also demonstrated novel adherence phenotypes with various strains of pathogenic Escherichia coli. This culture system will facilitate the study of inter-individual, functional studies of human intestinal epithelial cells, including host-microbial interactions.
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影响因子：
29.4
作者：
GRIFFIN, PM;OLMSTEAD, LC;PETRAS, RE
通讯作者：
PETRAS, RE
DOI：
10.1152/ajpgi.2001.280.5.g922
发表时间：
2001-05-01
影响因子：
4.5
作者：
Atuma, C;Strugala, V;Holm, L
通讯作者：
Holm, L
影响因子：
29.4
作者：
Darfeuille-Michaud, A;Neut, C;Colombel, JF
通讯作者：
Colombel, JF
影响因子：
64.8
作者：
Khor, Bernard;Gardet, Agnes;Xavier, Ramnik J.
通讯作者：
Xavier, Ramnik J.
影响因子：
29.7
作者：
Kaser A;Zeissig S;Blumberg RS
通讯作者：
Blumberg RS