A micropore array-based solid lift-off method for highly efficient and controllable cell alignment and spreading.
A micropore array-based solid lift-off method for highly efficient and controllable cell alignment and spreading.
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基于微孔阵列的固体剥离方法，用于高效、可控的细胞排列和铺展
DOI：
10.1038/s41378-020-00191-5
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发表时间：
2020
影响因子：
7.9
通讯作者：
Wang W
中科院分区：
文献类型：
--
作者：
Hun T;Liu Y;Guo Y;Sun Y;Fan Y;Wang W
解析细胞间以及细胞与微环境的相互作用，对于增进基础生物学知识和推动再生医学应用都至关重要。在以往研究中，细胞图案化已得到广泛探究。然而，已报道的方法无法同时实现对细胞排列以及细胞黏附/铺展的高效、高通量精确控制。在此，我们提出一种以微孔阵列作为荫罩的新型固态剥离方法。通过精心设计的荫罩，可同时实现对细胞排列以及细胞黏附/铺展的高效精确控制。该荫罩中央区域设有大微孔（捕获孔），周边区域设有小微孔（铺展孔），分别用于细胞捕获/排列控制和细胞黏附/铺展控制。固态剥离的作用过程如下：（1）蛋白质微图案通过捕获孔和铺展孔生成；（2）通过捕获孔实现细胞捕获/排列控制；（3）在荫罩剥离后，通过先前生成的蛋白质微图案控制细胞黏附/铺展。实现了高通量（2.4 - 3.2×10⁴个细胞/平方厘米）的细胞排列，且效率较高（单细胞、双细胞和三细胞排列的效率分别为86.2±3.2%、56.7±9.4%和51.1±4.0%）。利用小鼠骨骼肌成肌细胞，对细胞铺展的精确控制以及在调节细胞骨架和细胞间连接方面的应用进行了研究和验证。据我们所知，这是首次通过简单的固态剥离操作，同时展示高效且可控的多细胞排列以及细胞黏附/铺展的报告。本研究成功填补了文献空白，推动了细胞图案化在基础机制研究和应用医学领域有效且可重复的应用。
一种以精心设计的微孔阵列作为荫罩的固态剥离方法，能够实现对细胞图案化的高效精确控制。细胞图案化是基础生物学机制研究以及再生医学（如组织工程）应用技术开发的重要策略。在此，北京大学王巍教授带领的团队首次报告了通过简单的一步剥离操作，同时控制细胞排列和细胞黏附/铺展。其核心概念是，微孔阵列中央区域的大孔和周边区域的小孔分别控制细胞捕获与排列以及细胞黏附与铺展。他们展示了高通量、高效率的细胞排列以及对细胞铺展的精确控制。
Interpretation of cell–cell and cell-microenvironment interactions is critical for both advancing knowledge of basic biology and promoting applications of regenerative medicine. Cell patterning has been widely investigated in previous studies. However, the reported methods cannot simultaneously realize precise control of cell alignment and adhesion/spreading with a high efficiency at a high throughput. Here, a novel solid lift-off method with a micropore array as a shadow mask was proposed. Efficient and precise control of cell alignment and adhesion/spreading are simultaneously achieved via an ingeniously designed shadow mask, which contains large micropores (capture pores) in central areas and small micropores (spreading pores) in surrounding areas contributing to capture/alignment and adhesion/spreading control, respectively. The solid lift-off functions as follows: (1) protein micropattern generates through both the capture and spreading pores, (2) cell capture/alignment control is realized through the capture pores, and (3) cell adhesion/spreading is controlled through previously generated protein micropatterns after lift-off of the shadow mask. High-throughput (2.4–3.2 × 104 cells/cm2) cell alignments were achieved with high efficiencies (86.2 ± 3.2%, 56.7 ± 9.4% and 51.1 ± 4.0% for single-cell, double-cell, and triple-cell alignments, respectively). Precise control of cell spreading and applications for regulating cell skeletons and cell–cell junctions were investigated and verified using murine skeletal muscle myoblasts. To the best of our knowledge, this is the first report to demonstrate highly efficient and controllable multicell alignment and adhesion/spreading simultaneously via a simple solid lift-off operation. This study successfully fills a gap in literatures and promotes the effective and reproducible application of cell patterning in the fields of both basic mechanism studies and applied medicine. A solid lift-off method with an ingeniously designed micropore array as a shadow mask enables the efficient and precise control of cell patterning. Cell patterning is an important strategy for both basic biological mechanism studies and applicable technology developments in regenerative medicine, such as tissue engineering. Here, a team led by Prof. Wei Wang from Peking University presents the first report of a simultaneous control of cell alignment and adhesion/spreading via an easy single-step lift-off operation. The key concept is that the micropore array contains the large pores in central areas and small pores in surrounding areas controlling cell capture and alignment, and cell adhesion and spreading, respectively. They demonstrated high-throughput, high-efficiency cell alignment along with a precise control of cell spreading.
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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
影响因子：
14
作者：
Chew, Sing Ylan;Mi, Ruifa;Leong, Kam W.
通讯作者：
Leong, Kam W.
DOI：
10.1016/s1357-2725(01)00180-7
发表时间：
2002-07-01
影响因子：
4
作者：
Clark, P;Dunn, GA;Peckham, M
通讯作者：
Peckham, M
DOI：
10.1073/pnas.0408954102
发表时间：
2005-01-25
影响因子：
11.1
作者：
Jiang, XY;Bruzewicz, DA;Whitesides, GM
通讯作者：
Whitesides, GM
影响因子：
9.5
作者：
Lee, Donghee;Yang, Sung
通讯作者：
Yang, Sung