Human 2D Crypt Model for Assaying Intestinal Stem Cell Proliferation and Differentiation.
Human 2D Crypt Model for Assaying Intestinal Stem Cell Proliferation and Differentiation.
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DOI：
10.1021/acs.analchem.2c00905
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发表时间：
2022-07-05
影响因子：
7.4
通讯作者：
Allbritton, Nancy L.
中科院分区：
文献类型：
--
作者：
Wang, Yuli;Sims, Christopher E.;Allbritton, Nancy L.
肠是临床试验中药物不良反应的常见部位，因此寻求用于药物化合物的临床前筛选的改进的体外模型。一个平面的，自我更新的人肠单层平台的基础上原代成人胃肠干细胞，称为二维隐窝模型，已被开发用于筛选各种化合物对肠上皮细胞的影响。2D隐窝平台基于标准12孔板格式，由细胞培养插入物组成，胶原膜覆盖在图案化有微米级孔阵列的不可渗透膜上。这种2室格式能够应用生长因子的梯度，使得组织自组织成空间分离的干细胞和分化的细胞区室。图案化单层模仿肠上皮细胞，具有干细胞龛，迁移增殖和分化细胞。一旦建立，2D隐窝复制体内生理学的许多方面，包括细胞迁移、成熟和凋亡性细胞死亡。该系统的平面几何形状简化了测定期间的给药、取样和成像。建立了基于免疫荧光的测定以定量评估细胞密度、增殖、迁移、活力以及有丝分裂后谱系的丰度和定位作为时间的函数。该模型用于对组织稳态进行小规模的化合物筛选，包括信号分子、内源性激素/细胞因子和微生物代谢物。容易鉴定显著影响增殖和/或分化的命中化合物。2D隐窝平台代表了微生理系统开发的重大创新，用于模拟肠道上皮细胞进行化合物筛选。
Intestine is a common site of adverse drug effects in clinical trials, thus improved in vitro models for preclinical screening of pharmaceutical compounds are sought. A planar, self-renewing human intestinal monolayer platform based on primary adult gastrointestinal stem cells, termed the 2D crypt model, has been developed to screen for the effects of various compounds on the intestinal epithelium. The 2D crypt platform is a based on a standard 12-well plate format and consists of cell culture inserts with a collagen film overlaying an impermeable film patterned with an array of micron-scale holes. This 2-chamber format enables a gradient of growth factors to be applied such that the tissue self-organizes into spatially segregated stem and differentiated cell compartments. The patterned monolayer mimics a gut epithelium in possessing a stem-cell niche, migrating proliferative and differentiated cells. Once established the 2D crypts replicate many aspects of in vivo physiology, including cell migration, maturation and apoptotic cell death. The planar geometry of the system simplifies dosing, sampling and imaging during assay. An immunofluorescence-based assay was established to quantitatively assess cell density, proliferation, migration, viability, and the abundance and localization of post-mitotic lineages as a function of time. The model was used to perform a small-scale screen of compounds, including signaling molecules, endogenous hormones/cytokines, and microbial metabolites, on tissue homeostasis. Hit compounds that significantly impacted proliferation and/or differentiation were readily identified. The 2D crypt platform represents a significant innovation in the development of microphysiological systems for emulating the gut epithelium for compound screens.
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影响因子：
14.8
作者：
Hinman SS;Wang Y;Kim R;Allbritton NL
通讯作者：
Allbritton NL
影响因子：
64.5
作者：
Kaiko GE;Ryu SH;Koues OI;Collins PL;Solnica-Krezel L;Pearce EJ;Pearce EL;Oltz EM;Stappenbeck TS
通讯作者：
Stappenbeck TS
影响因子：
24.5
作者：
VanDussen KL;Marinshaw JM;Shaikh N;Miyoshi H;Moon C;Tarr PI;Ciorba MA;Stappenbeck TS
通讯作者：
Stappenbeck TS
影响因子：
6.4
作者：
GAMET, L;DAVIAUD, D;MURAT, JC
通讯作者：
MURAT, JC
影响因子：
4.6
作者：
VanDussen, Kelli L.;Carulli, Alexis J.;Samuelson, Linda C.
通讯作者：
Samuelson, Linda C.