Real-Time Microfluidic System for Studying Mammalian Cells in 3D Microenvironments

Lii, Jerry; Hsu, Wern-Jir; Parsa, Hesam; Das, Anshu; Rouse, Robert V.; Sia, Samuel K.

doi:10.1021/ac8000034

Cited by 91 publications

(74 citation statements)

References 36 publications

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“…The orientation of MDA-MB-231 cells inside the ECM was quantified by ImageJ and then displayed as population distributions of the measured angles. Because cell nucleus shape mostly represents its morphological change (33)(34)(35), the MDA-MB-231 cells were stained with Hoechst so that their nuclei would be visible under the confocal microscopy. Fig.…”

Section: Resultsmentioning

confidence: 99%

Oriented collagen fibers direct tumor cell intravasation

Han

Chen

Yuan

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

320

248

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In this work, we constructed a Collagen I-Matrigel composite extracellular matrix (ECM). The composite ECM was used to determine the influence of the local collagen fiber orientation on the collective intravasation ability of tumor cells. We found that the local fiber alignment enhanced cell-ECM interactions. Specifically, metastatic MDA-MB-231 breast cancer cells followed the local fiber alignment direction during the intravasation into rigid Matrigel (∼10 mg/mL protein concentration).M etastasis is a lethal milestone in cancer: Cells escape from the confinement of primary tumor sites (intravasation), invade tissues as well as the lymphatic and vascular systems, and finally colonize (extravasation) distant sites. It has been estimated that less than 1% of tumor cells undergo this process, but metastasis contributes to more than 90% of cancerrelated deaths (1, 2). Metastasis involves both genetic and epigenetic alternation of tumor cells, as well as external biochemical and biophysical microenvironments (3-5). Pathology studies suggest that metastatic tumor cells exhibit highly branched morphologies and distinct aligned registration with aligned extracellular matrix (ECM) during metastatic tumor progression (4, 5).We address three important questions concerning metastasis. (i) Can we build in vitro complex ECM structures with heterogeneously oriented collagen fibers and basement membrane components to mimic the cancer cell intravasation process? (ii) How does aligned collagen influence cell intravasation into/ through the basement membrane before entering vessels? (iii) After cell detachment from the primary tumor site, how does a heterogeneous ECM with a varying degree of local fiber alignment influence cell intravasation and subsequent penetration into the basement membrane during their intravasation process? The major obstacle to addressing these questions is the difficulty in constructing both an in vitro 3D microenvironment to mimic the above process and flexible controls of the environmental parameters, such as fiber orientations in a complex collagen/Matrigel composite, nutrition, oxygen, drug concentrations, etc.In breast cancer metastasis, cancer cells are believed to reorganize and progress through the interstitial ECM matrix, break through the basement membrane, and enter blood vessels or lymphatic capillaries (6-10). Fig. 1C presents a schematic illustration of the intravasation process in metastasis. Tumor-associated collagen signatures (TACS), basically environmentally elevated collagen density and collagen fiber reorganization, are used to stage mammary carcinoma tumor progression levels (6, 11-13). Fig. 1 presents hematoxylin/eosin (H&E)-stained biopsy slices of breast cancer imaged by second harmonic generation (SHG) under a two-photon confocal microscopy (A1R MP; Nikon) (detailed information provided in SI Appendix, SI Text) (6, 14, 15). Fig. 1 A, 1-3 shows the stained human invasive ductal carcinoma tumor at grade I. In the enlarged figures (Fig. 1 A, 2 and 3), the cells have well-defined bord...

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Section: Resultsmentioning

confidence: 99%

Oriented collagen fibers direct tumor cell intravasation

Han

Chen

Yuan

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

320

248

View full text Add to dashboard Cite

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“…To address the second hurdle, microfluidic systems and flow cytometry systems capable of probing deep within three-dimensional tissues in a high throughput manner are being developed. 17,[36][37][38] That ECM components can drive heterologous differentiation of stem cells signals a great opportunity for emerging fields of regenerative medicine. One could envision biomaterial and synthetic constructs tailored with specific ECMderived peptides to induce lineage-specific differentiation.…”

Section: Santiago Et Almentioning

confidence: 99%

Heterogeneous Differentiation of Human Mesenchymal Stem Cells in Response to Extended Culture in Extracellular Matrices

Santiago

Pogemiller

Ogle

2009

Tissue Engineering Part A

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Extracellular matrix proteins (ECMs) guide differentiation of adult stem cells, but the temporal distribution of differentiation (i.e., heterogeneity) in a given population has not been investigated. We tested the effect of individual ECM proteins on lineage commitment of human bone marrow-derived mesenchymal stem cells (MSCs) over time. We exposed stem cell populations to ECM proteins representing the primary tissue structures of the body (i.e., collagens type I, III, IV; laminin; and fibronectin) and determined the lineage commitment of the stem cells at 1, 7, and 14 days. We found that collagens that can participate in the formation of fibrils guide differentiation of cardiomyocytes, adipocytes, and osteoblasts. ECMs of the basement membrane initiate differentiation of cardiomyocytes and osteoblasts but not adipocytes, and small facilitator ECMs (e.g., fibronectin) do not significantly affect stem cell differentiation. Differentiation was ECM-dependent because culture on tissue culture polystyrene, with consistent cell morphology, proliferation, and death, initiated differentiation of osteoblasts only. Thus, we show that ECMs independently trigger differentiation of human adult MSCs and that differentiation in this context can be guided down multiple lineages using the same ECM stimulus. This work highlights the importance of more clearly defining progenitor populations, especially those cultured in the presence of ECMs before transplantation.

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“…In combination with traditional fluorescence microscopy, these new technologies will aid the study of subjects as far reaching as cell -cell communication, the role of the microenvironment in stem cell differentiation (Lii et al 2008), stem cell lineage commitment (Takano et al 2004), cancer metastasis and nerve regeneration. Optical injection adds value to this miniaturization trend by enabling a simple and convenient method to 'point-and-inject' material into a cell observed under the microscope.…”

Section: Optical Transfection With Fs-pulsed Sources: Targeted Singlementioning

confidence: 99%

Single cell optical transfection

Stevenson

Gunn‐Moore

Campbell

et al. 2010

J. R. Soc. Interface.

155

167

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The plasma membrane of a eukaryotic cell is impermeable to most hydrophilic substances, yet the insertion of these materials into cells is an extremely important and universal requirement for the cell biologist. To address this need, many transfection techniques have been developed including viral, lipoplex, polyplex, capillary microinjection, gene gun and electroporation. The current discussion explores a procedure called optical injection, where a laser field transiently increases the membrane permeability to allow species to be internalized. If the internalized substance is a nucleic acid, such as DNA, RNA or small interfering RNA (siRNA), then the process is called optical transfection. This contactless, aseptic, single cell transfection method provides a key nanosurgical tool to the microscopist-the intracellular delivery of reagents and single nanoscopic objects. The experimental possibilities enabled by this technology are only beginning to be realized. A review of optical transfection is presented, along with a forecast of future applications of this rapidly developing and exciting technology.

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Real-Time Microfluidic System for Studying Mammalian Cells in 3D Microenvironments

Cited by 91 publications

References 36 publications

Oriented collagen fibers direct tumor cell intravasation

Oriented collagen fibers direct tumor cell intravasation

Heterogeneous Differentiation of Human Mesenchymal Stem Cells in Response to Extended Culture in Extracellular Matrices

Single cell optical transfection

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