Quantification of nematic cell polarity in three-dimensional tissues

How epithelial cells coordinate their polarity to form functional tissues is an open question in cell biology. Here, we characterize a unique type of polarity found in liver tissue, nematic cell polarity, which is different from vectorial cell polarity in simple, sheet-like epithelia. We propose a conceptual and algorithmic framework to characterize complex patterns of polarity proteins on the surface of a cell in terms of a multipole expansion. To rigorously quantify previously observed tissue-level patterns of nematic cell polarity (Morales-Navarrete et al., eLife 2019), we introduce the concept of co-orientational order parameters, which generalize the known biaxial order parameters of the theory of liquid crystals. Applying these concepts to three-dimensional reconstructions of single cells from high-resolution imaging data of mouse liver tissue, we show that the axes of nematic cell polarity of hepatocytes exhibit local coordination and are aligned with the biaxially anisotropic sinusoidal network for blood transport. Our study characterizes liver tissue as a biological example of a biaxial liquid crystal. The general methodology developed here could be applied to other tissues and in-vitro organoids.

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Application of 3D printed model for planning the endoscopic endonasal transsphenoidal surgery

Scientific Reports ◽

10.1038/s41598-021-84779-5 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Xing Huang ◽

Ni Fan ◽

Hai-jun Wang ◽

Yan Zhou ◽

Xudong Li ◽

...

Keyword(s):

3D Printing ◽

Skull Base ◽

Transsphenoidal Surgery ◽

Three Dimensional ◽

Spatial Relationship ◽

Cranial Computed Tomography ◽

Imaging Data ◽

Endoscopic Endonasal ◽

Endoscopic Endonasal Transsphenoidal Surgery

AbstractThe application of 3D printing in planning endoscopic endonasal transsphenoidal surgery is illustrated based on the analysis of patients with intracranial skull base diseases who received treatment in our department. Cranial computed tomography/magnetic resonance imaging data are attained preoperatively, and three-dimensional reconstruction is performed using MIMICS (Materialise, Leuven, Belgium). Models of intracranial skull base diseases are printed using a 3D printer before surgery. The models clearly demonstrate the morphologies of the intracranial skull base diseases and the spatial relationship with adjacent large vessels and bones. The printing time of each model is 12.52–15.32 h, and the cost ranges from 900 to 1500 RMB. The operative approach was planned in vitro, and patients recovered postoperatively well without severe complications or death. In a questionnaire about the application of 3D printing, experienced neurosurgeons achieved scores of 7.8–8.8 out of 10, while unexperienced neurosurgeons achieved scores of 9.2–9.8. Resection of intracranial skull base lesions is demonstrated to be well assisted by 3D printing technique, which has great potential in disclosing adjacent anatomical relationships and providing the required help to clinical doctors in preoperative planning.

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A three-dimensional cell biology model of human hepatocellular carcinoma in vitro

Tumor Biology ◽

10.1007/s13277-010-0140-7 ◽

2010 ◽

Vol 32 (3) ◽

pp. 469-479 ◽

Cited By ~ 15

Author(s):

Jianhua Tang ◽

Jiefeng Cui ◽

Rongxin Chen ◽

Kun Guo ◽

Xiaonan Kang ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Cell Biology ◽

Three Dimensional ◽

Human Hepatocellular Carcinoma ◽

Dimensional Cell

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Epithelial Cell Chirality Revealed by Three-Dimensional Spontaneous Rotation

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1805932115 ◽

2018 ◽

Vol 115 (48) ◽

pp. 12188-12193 ◽

Cited By ~ 15

Author(s):

Amanda S. Chin ◽

Kathryn E. Worley ◽

Poulomi Ray ◽

Gurleen Kaur ◽

Jie Fan ◽

...

Keyword(s):

Environmental Factors ◽

Cell Polarity ◽

Three Dimensional ◽

Intrinsic Property ◽

Cell Aggregates ◽

Mechanistic Studies ◽

Directional Bias ◽

Self Organized ◽

Dependent Cell

Our understanding of the left–right (LR) asymmetry of embryonic development, in particular the contribution of intrinsic handedness of the cell or cell chirality, is limited due to the confounding systematic and environmental factors during morphogenesis and a ack of physiologically relevant in vitro 3D platforms. Here we report an efficient two-layered biomaterial platform for determining the chirality of individual cells, cell aggregates, and self-organized hollow epithelial spheroids. This bioengineered niche provides a uniform defined axis allowing for cells to rotate spontaneously with a directional bias toward either clockwise or counterclockwise directions. Mechanistic studies reveal an actin-dependent, cell-intrinsic property of 3D chirality that can be mediated by actin cross-linking via α-actinin-1. Our findings suggest that the gradient of extracellular matrix is an important biophysicochemical cue influencing cell polarity and chirality. Engineered biomaterial systems can serve as an effective platform for studying developmental asymmetry and screening for environmental factors causing birth defects.

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Planar cell polarity: moving from single cells to tissue-scale biology

Development ◽

10.1242/dev.186346 ◽

2020 ◽

Vol 147 (24) ◽

pp. dev186346

Author(s):

Marek Mlodzik

Keyword(s):

Cell Polarity ◽

Cell Biology ◽

Planar Cell Polarity ◽

Single Cells ◽

Field Studies ◽

Model System ◽

Convergent Extension ◽

Organ Function ◽

Biophysical Studies ◽

Cell Positioning

ABSTRACTPlanar cell polarity (PCP) reflects cellular orientation within the plane of an epithelium. PCP is crucial during many biological patterning processes and for organ function. It is omnipresent, from convergent-extension mechanisms during early development through to terminal organogenesis, and it regulates many aspects of cell positioning and orientation during tissue morphogenesis, organ development and homeostasis. Suzanne Eaton used the power of Drosophila as a model system to study PCP, but her vision of, and impact on, PCP studies in flies translates to all animal models. As I highlight here, Suzanne's incorporation of quantitative biophysical studies of whole tissues, integrated with the detailed cell biology of PCP phenomena, completely changed how the field studies this intriguing feature. Moreover, Suzanne's impact on ongoing and future PCP studies is fundamental, long-lasting and transformative.

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Spheroids as a Type of Three-Dimensional Cell Cultures—Examples of Methods of Preparation and the Most Important Application

International Journal of Molecular Sciences ◽

10.3390/ijms21176225 ◽

2020 ◽

Vol 21 (17) ◽

pp. 6225 ◽

Cited By ~ 1

Author(s):

Kamila Białkowska ◽

Piotr Komorowski ◽

Maria Bryszewska ◽

Katarzyna Miłowska

Keyword(s):

Cell Cultures ◽

Cell Biology ◽

Disease Modeling ◽

Three Dimensional ◽

3D Culture ◽

Matrix Interactions ◽

Vitro Model ◽

Extracellular Matrix Interactions ◽

Natural Cell

Cell cultures are very important for testing materials and drugs, and in the examination of cell biology and special cell mechanisms. The most popular models of cell culture are two-dimensional (2D) as monolayers, but this does not mimic the natural cell environment. Cells are mostly deprived of cell–cell and cell–extracellular matrix interactions. A much better in vitro model is three-dimensional (3D) culture. Because many cell lines have the ability to self-assemble, one 3D culturing method is to produce spheroids. There are several systems for culturing cells in spheroids, e.g., hanging drop, scaffolds and hydrogels, and these cultures have their applications in drug and nanoparticles testing, and disease modeling. In this paper we would like to present methods of preparation of spheroids in general and emphasize the most important applications.

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LKB1 kinase-dependent and -independent defects disrupt polarity and adhesion signaling to drive collagen remodeling during invasion

Molecular Biology of the Cell ◽

10.1091/mbc.e15-08-0569 ◽

2016 ◽

Vol 27 (7) ◽

pp. 1069-1084 ◽

Cited By ~ 14

Author(s):

Jessica Konen ◽

Scott Wilkinson ◽

Byoungkoo Lee ◽

Haian Fu ◽

Wei Zhou ◽

...

Keyword(s):

Cell Polarity ◽

Cancer Metastasis ◽

Kinase Activity ◽

Rho Gtpase ◽

Three Dimensional ◽

Serine Threonine Kinase ◽

Collagen Remodeling ◽

Threonine Kinase ◽

Three Dimensional Models

LKB1 is a serine/threonine kinase and a commonly mutated gene in lung adenocarcinoma. The majority of LKB1 mutations are truncations that disrupt its kinase activity and remove its C-terminal domain (CTD). Because LKB1 inactivation drives cancer metastasis in mice and leads to aberrant cell invasion in vitro, we sought to determine how compromised LKB1 function affects lung cancer cell polarity and invasion. Using three-dimensional models, we show that LKB1 kinase activity is essential for focal adhesion kinase–mediated cell adhesion and subsequent collagen remodeling but not cell polarity. Instead, cell polarity is overseen by the kinase-independent function of its CTD and more specifically its farnesylation. This occurs through a mesenchymal-amoeboid morphological switch that signals through the Rho-GTPase RhoA. These data suggest that a combination of kinase-dependent and -independent defects by LKB1 inactivation creates a uniquely invasive cell with aberrant polarity and adhesion signaling that drives invasion into the microenvironment.

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Feasibility of Biomedical Applications of Hall Effect Imaging

Ultrasonic Imaging ◽

10.1177/016173460002200204 ◽

2000 ◽

Vol 22 (2) ◽

pp. 123-136 ◽

Cited By ~ 23

Author(s):

Han Wen

Keyword(s):

Hall Effect ◽

Biomedical Applications ◽

Three Dimensional ◽

Imaging Data ◽

Theoretical Understanding ◽

Complex Samples ◽

Quantitative Measurements ◽

Basic Physics ◽

A New Technique

Hall effect imaging is a new technique for mapping the electrical properties of a sample. Its principle has been demonstrated in two- and three-dimensional phantom images. Based on the experimental data and theoretical understanding of this technique developed over the past few years, this paper addresses the most relevant question for biomedical applications: whether Hall effect imaging is ultimately applicable to complex biological systems such as the human body. The arguments are given at the basic physics level, so that the conclusion is independent of current technology status. These arguments are corroborated with imaging data of an aorta sample. The conclusion is that Hall effect imaging is not suited for quantifying the electrical constants in complex biological samples. This technique is able to produce high-resolution volume images of samples in vitro that reflect their electrical heterogeneity. However, quantitative measurements of electrical constants are not practical for complex samples.

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Vinculin regulates directionality and cell polarity in two- and three-dimensional matrix and three-dimensional microtrack migration

Molecular Biology of the Cell ◽

10.1091/mbc.e15-06-0432 ◽

2016 ◽

Vol 27 (9) ◽

pp. 1431-1441 ◽

Cited By ~ 35

Author(s):

Aniqua Rahman ◽

Shawn P. Carey ◽

Casey M. Kraning-Rush ◽

Zachary E. Goldblatt ◽

Francois Bordeleau ◽

...

Keyword(s):

Cell Polarity ◽

Focal Adhesion ◽

Molecular Mechanisms ◽

Three Dimensional ◽

Focal Adhesions ◽

Three Dimensions ◽

Collagen Matrices ◽

Migration Direction ◽

Unidirectional Motion

During metastasis, cells can use proteolytic activity to form tube-like “microtracks” within the extracellular matrix (ECM). Using these microtracks, cells can migrate unimpeded through the stroma. To investigate the molecular mechanisms of microtrack migration, we developed an in vitro three-dimensional (3D) micromolded collagen platform. When in microtracks, cells tend to migrate unidirectionally. Because focal adhesions are the primary mechanism by which cells interact with the ECM, we examined the roles of several focal adhesion molecules in driving unidirectional motion. Vinculin knockdown results in the repeated reversal of migration direction compared with control cells. Tracking the position of the Golgi centroid relative to the position of the nucleus centroid reveals that vinculin knockdown disrupts cell polarity in microtracks. Vinculin also directs migration on two-dimensional (2D) substrates and in 3D uniform collagen matrices, as indicated by reduced speed, shorter net displacement, and decreased directionality in vinculin-deficient cells. In addition, vinculin is necessary for focal adhesion kinase (FAK) activation in three dimensions, as vinculin knockdown results in reduced FAK activation in both 3D uniform collagen matrices and microtracks but not on 2D substrates, and, accordingly, FAK inhibition halts cell migration in 3D microtracks. Together these data indicate that vinculin plays a key role in polarization during migration.

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Optical Cellular Micromotion: A New Paradigm to Measure Tumour Cells Invasion in 3D Tumour Environments

10.1101/2021.08.26.457857 ◽

2021 ◽

Author(s):

Zhaobin Guo ◽

Chih-Tsung Yang ◽

Chia-Chi Chien ◽

Luke Selth ◽

Pierre Bagnaninchi ◽

...

Keyword(s):

Single Cell ◽

Cell Invasion ◽

Tumour Cell ◽

Single Cells ◽

Three Dimensional ◽

Tumour Cells ◽

Digital Holographic Microscopy ◽

New Paradigm ◽

Cell Invasiveness

Measuring tumour cell invasiveness through three-dimensional (3D) tissues, particularly at the single cell level, can provide important mechanistic understanding and assist in identifying therapeutic targets of tumour invasion. However, current experimental approaches, including standard in vitro invasion assays, have limited physiological relevance and offer insufficient insight about the vast heterogeneity in tumour cell migration through tissues. To address these issues, here we report on the concept of optical cellular micromotion, where digital holographic microscopy (DHM) is used to map the optical thickness fluctuations at sub-micron scale within single cells. These fluctuations are driven by the dynamic movement of subcellular structures including the cytoskeleton and inherently associated with the biological processes involved in cell invasion within tissues. We experimentally demonstrate that the optical cellular micromotion correlates with tumour cells motility and invasiveness both at the population and single cell levels. In addition, the optical cellular micromotion significantly reduced upon treatment with migrastatic drugs that inhibit tumour cell invasion. These results demonstrate that micromotion measurements can rapidly and non-invasively determine the invasive behaviour of single tumour cells within tissues, yielding a new and powerful tool to assess the efficacy of approaches targeting tumour cell invasiveness.

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In Vitro Tissue Microarrays for Quick and Efficient Spheroid Characterization

SLAS DISCOVERY Advancing Life Sciences ◽

10.1177/2472555217740576 ◽

2017 ◽

Vol 23 (2) ◽

pp. 211-217 ◽

Cited By ~ 5

Author(s):

D. P. Ivanov ◽

A. M. Grabowska

Keyword(s):

Single Cells ◽

Three Dimensional ◽

Tissue Microarrays ◽

3D Models ◽

Morphological Examination ◽

Protein Targets ◽

Cell Monolayers ◽

Her 2 ◽

Models Of Disease

Three-dimensional (3D) in vitro microphysiological cultures, such as spheroids and organoids, promise increased patient relevance and therapeutic predictivity compared with reductionist cell monolayers. However, high-throughput characterization techniques for 3D models are currently limited to simplistic live/dead assays. By sectioning and staining in vitro microtissues, researchers can examine their structure; detect DNA, RNA, and protein targets; and visualize them at the level of single cells. The morphological examination and immunochemistry staining for in vitro cultures has historically been done in a laborious manner involving testing one set of cultures at a time. We have developed a technology to rapidly screen spheroid phenotype and protein expression by arranging 66 spheroids in a gel array for paraffin embedding, sectioning, and immunohistochemsitry. The process is quick, mostly automatable, and uses 11 times less reagents than conventional techniques. Here we showcase the capabilities of the technique in an array made up of 11 different cell lines stained in conventional hematoxylin and eosin (H&E) staining, as well as immunohistochemistry staining for estrogen (ER), progesterone (PR), and human epidermal growth factor (Her-2) receptors, and TP53. This new methodology can be used in optimizing stem cell–based models of disease and development, for tissue engineering, safety screening, and efficacy screens in cancer research.

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