Endogenous Optical Signals Reveal Changes of Elastin and Collagen Organization During Differentiation of Mouse Embryonic Stem Cells.
Endogenous Optical Signals Reveal Changes of Elastin and Collagen Organization During Differentiation of Mouse Embryonic Stem Cells.
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内源性光信号揭示小鼠胚胎干细胞分化过程中弹性蛋白和胶原蛋白组织的变化。
DOI：
10.1089/ten.tec.2014.0699
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发表时间：
2015
期刊：
影响因子：
--
通讯作者：
Ogle,BrendaM
中科院分区：
文献类型：
--
作者：
Thimm,TerraN;Squirrell,JayneM;Liu,Yuming;Eliceiri,KevinW;Ogle,BrendaM
细胞外基质成分（ECM）最近被证明影响干细胞规格。然而，评估干细胞- ecm相互作用的时空动态一直具有挑战性，因为迄今为止使用的大多数方法都需要破坏或固定样本。我们研究了在小鼠胚胎干细胞分化过程中利用两种重要ECM蛋白的内源性光信号的效果，弹性蛋白（Eln）通过自身荧光，I型胶原（ColI）通过二次谐波产生（SHG）。在发现抗体标记与Eln的内源性荧光信号之间存在良好的重叠后，我们利用这一内源性荧光信号绘制了小鼠胚状体分化过程中Eln和ColI的时间变化，发现Eln在第9天呈上升趋势，到第12天略有下降，而Col1在12天内稳步上升。此外，我们将内源性荧光成像和SHG与心肌细胞抗体标记相结合，研究Eln和ColI积累与心肌细胞分化之间的空间关系。Eln普遍存在，在心肌细胞分化区富集，而ColI与心肌细胞分化呈负相关。这项工作为利用内源性光信号提供了重要的第一步，这些信号可以在活细胞中可视化，以了解ECM与心肌细胞发育之间的关系，并为未来干细胞-ECM相互作用和与干细胞以及其他细胞和组织类型相关的动力学研究奠定了基础。
Components of the extracellular matrix (ECM) have recently been shown to influence stem cell specification. However, it has been challenging to assess the spatial and temporal dynamics of stem cell-ECM interactions because most methodologies utilized to date require sample destruction or fixation. We examined the efficacy of utilizing the endogenous optical signals of two important ECM proteins, elastin (Eln), through autofluorescence, and type I collagen (ColI), through second harmonic generation (SHG), during mouse embryonic stem cell differentiation. After finding favorable overlap between antibody labeling and the endogenous fluorescent signal of Eln, we used this endogenous signal to map temporal changes in Eln and ColI during murine embryoid body differentiation and found that Eln increases until day 9 and then decreases slightly by day 12, while Col1 steadily increases over the 12-day period. Furthermore, we combined endogenous fluorescence imaging and SHG with antibody labeling of cardiomyocytes to examine the spatial relationship between Eln and ColI accumulation and cardiomyocyte differentiation. Eln was ubiquitously present, with enrichment in regions with cardiomyocyte differentiation, while there was an inverse correlation between ColI and cardiomyocyte differentiation. This work provides an important first step for utilizing endogenous optical signals, which can be visualized in living cells, to understand the relationship between the ECM and cardiomyocyte development and sets the stage for future studies of stem cell-ECM interactions and dynamics relevant to stem cells as well as other cell and tissue types.
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影响因子：
37.8
作者：
Go AS;Mozaffarian D;Roger VL;Benjamin EJ;Berry JD;Blaha MJ;Dai S;Ford ES;Fox CS;Franco S;Fullerton HJ;Gillespie C;Hailpern SM;Heit JA;Howard VJ;Huffman MD;Judd SE;Kissela BM;Kittner SJ;Lackland DT;Lichtman JH;Lisabeth LD;Mackey RH;Magid DJ;Marcus GM;Marelli A;Matchar DB;McGuire DK;Mohler ER 3rd;Moy CS;Mussolino ME;Neumar RW;Nichol G;Pandey DK;Paynter NP;Reeves MJ;Sorlie PD;Stein J;Towfighi A;Turan TN;Virani SS;Wong ND;Woo D;Turner MB;American Heart Association Statistics Committee and Stroke Statistics Subcommittee
通讯作者：
American Heart Association Statistics Committee and Stroke Statistics Subcommittee
影响因子：
3.8
作者：
Balestrini, Jenna L.;Niklason, Laura E.
通讯作者：
Niklason, Laura E.
影响因子：
3.6
作者：
Hall G;Tilbury KB;Campbell KR;Eliceiri KW;Campagnola PJ
通讯作者：
Campagnola PJ
DOI：
10.1007/978-1-4757-1287-2_19
发表时间：
1985
期刊：
Advances in myocardiology
影响因子：
--
作者：
Robinson,TF;Cohen-Gould,L;Remily,RM;Capasso,JM;Factor,SM
通讯作者：
Factor,SM
影响因子：
1
作者：
Jakob, Andre;Unger, Sheila;Stiller, Brigitte
通讯作者：
Stiller, Brigitte