Synthetic alternatives to Matrigel

Aisenbrey, Elizabeth A.; Murphy, William L.

doi:10.1038/s41578-020-0199-8

Cited by 643 publications

(613 citation statements)

References 191 publications

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“…Recent 3D models of trophoblast migration have used biomaterials as a matrix around invading trophoblast cells as a means of replicating trophoblast invasion through tissue (Buck et al, 2015;Chang et al, 2018;Kuo et al, 2016;Kuo et al, 2018;Kuo et al, 2019;Wang et al, 2013;Wang et al, 2012); however, many of these models often employ biomaterials such as Matrigel that have limitations. For example, Matrigel has been found to have significant batch-to-batch variations in biomechanical properties, extracellular matrix content, and protein content (Aisenbrey and Murphy, 2020;Hughes et al, 2010;Vukicevic et al, 1992). The GelMA hydrogel system used in this work offers a number of potential advantages specific for investigating trophoblast activity.…”

Section: Discussionmentioning

confidence: 99%

The role of pregnancy-specific glycoproteins on trophoblast motility in three-dimensional gelatin hydrogels

Zambuto

Rattila

Dveksler

et al. 2020

Preprint

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SUMMARYTrophoblast invasion is a complex biological process necessary for establishment of pregnancy; however, much remains unknown regarding what signaling factors coordinate the extent of invasion. Pregnancy-specific glycoproteins (PSGs) are some of the most abundant circulating trophoblastic proteins in maternal blood during human pregnancy, with maternal serum concentrations rising to as high as 200-400 μg/mL at term. Here, we employ three-dimensional (3D) trophoblast motility assays consisting of trophoblast spheroids encapsulated in 3D gelatin hydrogels to quantify trophoblast outgrowth area, viability, and cytotoxicity in the presence of PSG1 and PSG9 as well as epidermal growth factor and Nodal. We show PSG9 reduces trophoblast motility whereas PSG1 increases motility. Further, we assess bulk nascent protein production by encapsulated spheroids to highlight the potential of this approach to assess trophoblast response (motility, remodeling) to soluble factors and extracellular matrix cues. Such models provide an important platform to develop a deeper understanding of early pregnancy.Graphical Abstract

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Section: Discussionmentioning

confidence: 99%

The role of pregnancy-specific glycoproteins on trophoblast motility in three-dimensional gelatin hydrogels

Zambuto

Rattila

Dveksler

et al. 2020

Preprint

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“…Tumor organoids can be isolated from experimental mouse models as well as human patient primary and metastatic tumors, allowing for the representation of a wide range of tumor phenotypes and genotypes. To observe and perturb complex interactions amongst tumor cells, normal cells, and stroma, multicellular tumor organoids are embedded in a variety of 3D matrix platforms [2,11,12,20,32,[43][44][45]. In this way, investigators have used tumor organoids in 3D culture to uncover essential aspects of tumor formation, met a s t a s i s , i n t e r c e l l u l a r t u m o r -t u m o r a n d t u m o rmicroenvironmental interactions, and therapeutic drug resistance [4-6, 10, 24, 46, 61].…”

Section: Introductionmentioning

confidence: 99%

Optimal, Large-Scale Propagation of Mouse Mammary Tumor Organoids

Wrenn

Moore

Greenwood

et al. 2020

J Mammary Gland Biol Neoplasia

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Tumor organoids mimic the architecture and heterogeneity of in vivo tumors and enable studies of collective interactions between tumor cells as well as with their surrounding microenvironment. Although tumor organoids hold significant promise as cancer models, they are also more costly and labor-intensive to cultivate than traditional 2D cell culture. We sought to identify critical factors regulating organoid growth ex vivo, and to use these observations to develop a more efficient organoid expansion method. Using time-lapse imaging of mouse mammary tumor organoids in 3D culture, we observed that outgrowth potential varies nonlinearly with initial organoid size. Maximal outgrowth occurred in organoids with a starting size between~10 to 1000 cells. Based on these observations, we developed a suspension culture method that maintains organoids in the ideal size range, enabling expansion from 1 million to over 100 million cells in less than 2 weeks and less than 3 hours of hands-on time. Our method facilitates the rapid, cost-effective expansion of organoids for CRISPR based studies and other assays requiring a large amount of organoid starting material.

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“…For example, Matrigel, which is reconstituted from the mouse sarcoma and composed of laminin, entactin, collagens and heparin sulfate proteoglycan plus an unknown mixture of growth factors and enzymes, is commonly used in biology. [5,6] It is important to understand that some properties of the hydrogels have major drawbacks, such as very low rigidity, which does not mimic the naturally stiff environment of some tissues. [7] To ensure optimal performance, swelling of these 3D biomaterials, their permeability to different molecules, interaction with media and the immobilization of biomolecules have to be optimized.…”

Section: Introductionmentioning

confidence: 99%

“…All these limitations make hydrogels unsuitable for tissue modeling in vitro. [6] Consequently, toward the development of scalable 3D culture system, cost-effective, chemically defined, and reproducible culture systems are required.…”

Section: Introductionmentioning

confidence: 99%

3D synthetic microscaffolds promote homogenous expression of NANOG in mouse embryonic stem cells

Bakhtina

Müller

Wischnewski

et al. 2020

Preprint

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The development of in vitro models, which accurately recapitulate early embryonic development, is one of the fundamental challenges in stem cell research. Most of the currently employed approaches involve the culture of embryonic stem cells (ESCs) on two-dimensional (2D) surfaces. However, the monolayer nature of these cultures does not permit cells to grow and proliferate in realistic three-dimensional (3D) microenvironments, as in an early embryo. In this paper, novel 3D synthetic scaffold arrays, fabricated by two-photon polymerization photolithography, are utilized to mimic tissue-specific architecture, enabling cell-to-matrix interaction and cell-to-cell communication in vitro. Mouse ESCs (mESCs) are able to grow and proliferate on these structures and maintain their pluripotent state. Furthermore, the 3D microscaffold arrays are integrated into a microscopy slide allowing the evaluation of the expression of key pluripotency factors at the single-cell level. Comparing 2D and 3D surfaces, mESCs grown in serum+LIF on 3D microscaffolds exhibit a stronger and more homogenous expression of NANOG and OCT4 pluripotency factors, than cells cultivated in 2i media, demonstrating that 3D microscaffolds capture naive pluripotency in vitro. Thus, the slide affords a novel and unique tool to model and study mammalian early development with greater physiological relevance than conventional 2D cultures.

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Synthetic alternatives to Matrigel

Cited by 643 publications

References 191 publications

The role of pregnancy-specific glycoproteins on trophoblast motility in three-dimensional gelatin hydrogels

The role of pregnancy-specific glycoproteins on trophoblast motility in three-dimensional gelatin hydrogels

Optimal, Large-Scale Propagation of Mouse Mammary Tumor Organoids

3D synthetic microscaffolds promote homogenous expression of NANOG in mouse embryonic stem cells

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