Single Cell Imaging to Probe Mesenchymal Stem Cell N-Cadherin Mediated Signaling within Hydrogels.
Single Cell Imaging to Probe Mesenchymal Stem Cell N-Cadherin Mediated Signaling within Hydrogels.
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DOI：
10.1007/s10439-016-1622-6
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发表时间：
2016-06
影响因子：
3.8
通讯作者：
Burdick JA
中科院分区：
文献类型：
--
作者：
Vega SL;Kwon M;Mauck RL;Burdick JA
N - 钙黏蛋白（Ncad）介导细胞间相互作用，调节β - 连环蛋白（βcat）信号传导，并促进间充质谱系细胞的软骨分化。在此，我们利用共聚焦成像研究了在促进软骨分化的条件下，Ncad相互作用对三维水凝胶环境中单个间充质干细胞（MSC）行为的影响。将人MSC光包封在经Ncad模拟肽功能化的透明质酸（HA）水凝胶中，并与含有对照非活性肽（Ctrl）的环境中的细胞进行比较。通过单细胞成像，我们观察到与Ctrl水凝胶相比，在Ncad水凝胶中培养3天后，细胞膜βcat、细胞核βcat以及细胞圆度均显著增加。在将细胞包封于Ncad水凝胶之前，通过用Ncad特异性抗体预处理，细胞膜和细胞核βcat的定位程度以及MSC圆度降低至Ctrl水凝胶水平，这证实了该肽的活性。有趣的是，在Ctrl水凝胶中的双细胞簇中，βcat细胞核定位显著增加（>80%），这比在Ncad水凝胶中的双细胞簇中βcat细胞核定位的增加（约30%）要大得多。总之，我们利用荧光成像证明了Ncad介导的单细胞对水凝胶内朝向软骨形成的发育信号的反应。
N-cadherin (Ncad) mediates cell-cell interactions, regulates β-catenin (βcat) signaling, and promotes the chondrogenic differentiation of mesenchymal lineage cells. Here, we utilized confocal imaging to investigate the influence of Ncad interactions on single mesenchymal stem cell (MSC) behavior within 3-dimensional hydrogel environments under conditions that promote chondrogenic differentiation. Human MSCs were photoencapsulated in hyaluronic acid (HA) hydrogels functionalized with Ncad mimetic peptides and compared to cells in environments with control non-active peptides (Ctrl). Using single-cell imaging, we observed a significant increase in membrane βcat, nuclear βcat, and cell roundness after 3 days in Ncad hydrogels compared to Ctrl hydrogels. The extent of membrane and nuclear βcat localization and MSC roundness decreased to Ctrl hydrogel levels via pre-treatment with Ncad-specific antibodies prior to encapsulation in the Ncad hydrogels, confirming the activity of the peptide. Interestingly, there was a pronounced (>80% increase) in βcat nuclear localization in two-cell clusters within the Ctrl hydrogels, which was much greater than the increase (~30%) in βcat nuclear localization in two-cell clusters within the Ncad hydrogels. In summary, we utilized fluorescent imaging to demonstrate Ncad-mediated single cell responses to developmental cues within hydrogels towards chondrogenesis.
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影响因子：
5.6
作者：
Lisignoli, G;Cristino, S;Facchini, A
通讯作者：
Facchini, A
影响因子：
2.4
作者：
Huang AH;Farrell MJ;Mauck RL
通讯作者：
Mauck RL
DOI：
10.1073/pnas.1214100110
发表时间：
2013-06-18
影响因子：
11.1
作者：
Bian, Liming;Guvendiren, Murat;Burdick, Jason A.
通讯作者：
Burdick, Jason A.
DOI：
10.1073/pnas.0903269107
发表时间：
2010-03-16
影响因子：
11.1
作者：
Kilian, Kristopher A.;Bugarija, Branimir;Mrksich, Milan
通讯作者：
Mrksich, Milan
DOI：
10.1073/pnas.95.26.15339
发表时间：
1998-12-22
影响因子：
11.1
作者：
Sadot, E;Simcha, I;Geiger, B
通讯作者：
Geiger, B