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Stem/Proliferative and Differentiated Cells within Primary Murine Colonic Epithelium Display Distinct Intracellular Free Ca2+ Signal Codes
Advanced Healthcare Materials ( IF 10.0 ) Pub Date : 2021-09-12 , DOI: 10.1002/adhm.202101318 Sebastian Mestril 1 , Raehyun Kim 2 , Samuel S Hinman 2 , Shawn M Gomez 1, 3 , Nancy L Allbritton 2
Advanced Healthcare Materials ( IF 10.0 ) Pub Date : 2021-09-12 , DOI: 10.1002/adhm.202101318 Sebastian Mestril 1 , Raehyun Kim 2 , Samuel S Hinman 2 , Shawn M Gomez 1, 3 , Nancy L Allbritton 2
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The second messenger, intracellular free calcium (Ca2+), acts to transduce mitogenic and differentiation signals incoming to the colonic epithelium. A self-renewing monolayer of primary murine colonic epithelial cells is formed over a soft, transparent hydrogel matrix for the scalable analysis of intracellular Ca2+ transients. Cultures that are enriched for stem/proliferative cells exhibit repetitive, high frequency (≈25 peaks h−1), and short pulse width (≈25 s) Ca2+ transients. Upon cell differentiation the transient frequency declines by 50% and pulse width widens by 200%. Metabolites and growth factors that are known to modulate stem cell proliferation and differentiation through Wnt and Notch signaling pathways, including CHIR-99021, N-[(3,5-Difluorophenyl)acetyl]-L-alanyl-2-phenylglycine-1,1-dimethylethyl ester (DAPT), bone morphogenetic proteins (BMPs), and butyrate, also modulate Ca2+ oscillation patterns in a consistent manner. Increasing the stiffness of the supportive matrix from 200 Pa to 3 GPa shifts Ca2+ transient patterns toward those resembling differentiated cells. The ability to monitor Ca2+ oscillations with the spatial and temporal resolution offered by this platform, combined with its amenability to high-content screens, provides a powerful tool for investigating real-time communication within a wide range of primary tissues in addition to the colonic epithelium.
中文翻译：
原代小鼠结肠上皮内的干细胞/增殖细胞和分化细胞显示出独特的细胞内游离 Ca2+ 信号代码
第二信使，细胞内游离钙 (Ca 2+ )，作用是转导传入结肠上皮的促有丝分裂和分化信号。在柔软、透明的水凝胶基质上形成自我更新的单层原代小鼠结肠上皮细胞，用于细胞内 Ca 2+瞬变的可扩展分析。富集干细胞/增殖细胞的培养物表现出重复的高频（约25个峰h -1 ）和短脉冲宽度（约25秒）Ca 2+瞬变。细胞分化后，瞬态频率下降 50%，脉冲宽度加宽 200%。已知通过 Wnt 和 Notch 信号通路调节干细胞增殖和分化的代谢物和生长因子，包括 CHIR-99021、N-[(3,5-二氟苯基)乙酰基]-L-丙氨酰-2-苯基甘氨酸-1,1 -二甲基乙酯 (DAPT)、骨形态发生蛋白 (BMP) 和丁酸盐也以一致的方式调节 Ca 2+振荡模式。将支撑基质的刚度从 200 Pa 增加到 3 GPa 会将 Ca 2+瞬态模式转变为类似于分化细胞的模式。该平台提供的空间和时间分辨率监测 Ca 2+振荡的能力，结合其对高内涵屏幕的适应性，为研究各种原代组织内的实时通信提供了强大的工具。结肠上皮。
更新日期：2021-11-17
中文翻译：
原代小鼠结肠上皮内的干细胞/增殖细胞和分化细胞显示出独特的细胞内游离 Ca2+ 信号代码
第二信使，细胞内游离钙 (Ca 2+ )，作用是转导传入结肠上皮的促有丝分裂和分化信号。在柔软、透明的水凝胶基质上形成自我更新的单层原代小鼠结肠上皮细胞，用于细胞内 Ca 2+瞬变的可扩展分析。富集干细胞/增殖细胞的培养物表现出重复的高频（约25个峰h -1 ）和短脉冲宽度（约25秒）Ca 2+瞬变。细胞分化后，瞬态频率下降 50%，脉冲宽度加宽 200%。已知通过 Wnt 和 Notch 信号通路调节干细胞增殖和分化的代谢物和生长因子，包括 CHIR-99021、N-[(3,5-二氟苯基)乙酰基]-L-丙氨酰-2-苯基甘氨酸-1,1 -二甲基乙酯 (DAPT)、骨形态发生蛋白 (BMP) 和丁酸盐也以一致的方式调节 Ca 2+振荡模式。将支撑基质的刚度从 200 Pa 增加到 3 GPa 会将 Ca 2+瞬态模式转变为类似于分化细胞的模式。该平台提供的空间和时间分辨率监测 Ca 2+振荡的能力，结合其对高内涵屏幕的适应性，为研究各种原代组织内的实时通信提供了强大的工具。结肠上皮。
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