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Article
| Open Access
ReLo is a simple and rapid colocalization assay to identify and characterize direct protein–protein interactions
Characterising interactions between proteins that are large and poorly soluble remains challenging. Here, the authors describe ReLo, a rapid and versatile eukaryotic cell culture-based method for detecting and studying direct interactions between structurally complex proteins.
- Harpreet Kaur Salgania
- , Jutta Metz
- & Mandy Jeske
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Article
| Open Access
Trophoblast stem cell-based organoid models of the human placental barrier
The placenta is a transient organ that regulates the fetal environment, but our understanding of placental barrier function has been hampered by the lack of in vitro models. Here they develop human placental organoids that resemble the placental villus and form an intact syncytiotrophoblast barrier when cultured in a column model.
- Takeshi Hori
- , Hiroaki Okae
- & Hirokazu Kaji
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| Open Access
Self-renewing human naïve pluripotent stem cells dedifferentiate in 3D culture and form blastoids spontaneously
Blastoids are emerging models for early embryo development exploration in vitro. Here, authors found self-renewing human naïve PSCs spontaneously and efficiently give rise to blastoids upon three-dimensional suspension culture.
- Mingyue Guo
- , Jinyi Wu
- & José C. R. Silva
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Article
| Open Access
Cellular model system to dissect the isoform-selectivity of Akt inhibitors
Elucidating specific effects of protein kinase Akt isoforms remains challenging. Here, the authors establish an Akt isoform-dependent cellular model system and use it, together with X-ray crystallography and structure-based ligand design, to develop isoform-selective covalent-allosteric Akt inhibitors
- Lena Quambusch
- , Laura Depta
- & Daniel Rauh
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Article
| Open Access
Exploiting oxidative phosphorylation to promote the stem and immunoevasive properties of pancreatic cancer stem cells
Long-term cultures of pancreatic cancer stem cells (PaCSCs) are difficult to obtain. Here, the authors present a 2D culture method, based on the use of galactose, to establish cell cultures from pancreatic ductal adenocarcinoma xenotransplants enriched in PaCSCs.
- Sandra Valle
- , Sonia Alcalá
- & Bruno Sainz Jr
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Article
| Open Access
Functional analysis of genetic variants in the high-risk breast cancer susceptibility gene PALB2
PALB2 is an established breast cancer risk gene but the pathogenicity of many variants remains uncharacterised. Here, the authors present a cDNA-based system for the functional analysis of PALB2 variants of unknown significance.
- Rick A. C. M. Boonen
- , Amélie Rodrigue
- & Haico van Attikum
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Article
| Open Access
Expansion of functional personalized cells with specific transgene combinations
Personalised medicine requires cell cultures from defined genetic backgrounds, but providing sufficient numbers of cells is a challenge. Here the authors develop gene cocktails to expand primary cells from a variety of different tissues and species, and show that expanded endothelial and hepatic cells retain properties of the differentiated phenotype.
- Christoph Lipps
- , Franziska Klein
- & Tobias May
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Article
| Open Access
High-efficiency RNA-based reprogramming of human primary fibroblasts
Induced pluripotent stem cells (iPSCs) have potential for regenerative medicine applications, but are generated with very low efficiency. Here, the authors show highly efficient reprogramming of human primary fibroblasts to iPSCs via the synergistic activity of synthetic modified mRNAs, mature miRNA mimics, and optimized culture methods.
- Igor Kogut
- , Sandra M. McCarthy
- & Ganna Bilousova
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Article
| Open Access
3D microniches reveal the importance of cell size and shape
Little is known about how geometric cues affect cell function and gene expression in 3D settings. Here the authors use microniches of different geometries to control cell volume and shape, and by extension cell phenotype and lineage.
- Min Bao
- , Jing Xie
- & Wilhelm T. S. Huck
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Article
| Open Access
Multiscale cytometry and regulation of 3D cell cultures on a chip
3D cell culture is more relevant than the two-dimensional format, but methods for parallel analysis and temporal regulation of the microenvironment are limited. Here the authors develop a droplet microfluidics system to perform long-term culture of 3D spheroids, enabling multiscale cytometry of individual cells within the spheroid.
- Sébastien Sart
- , Raphaël F.-X. Tomasi
- & Charles N. Baroud
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Article
| Open Access
Human serum-derived protein removes the need for coating in defined human pluripotent stem cell culture
Improved culture methods are needed to reliably grow human pluripotent stem cells (hPSCs) on a large scale. Here, the authors identify a xeno-free medium with a supplement of Inter-α-inhibitor that supports long-term propagation and improved single-cell passaging of hPSCs on uncoated plastic.
- Sara Pijuan-Galitó
- , Christoffer Tamm
- & Cecilia Annerén
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Article
| Open Access
Characterization of twenty-five ovarian tumour cell lines that phenocopy primary tumours
Despite methodological improvements, it remains extremely difficult to routinely establish cell lines from human primary tumours. Here, the authors develop a culture medium that allows efficient cell line establishment and use this to generate 25 ovarian cancer cell lines.
- Tan A. Ince
- , Aurea D. Sousa
- & Gordon B. Mills
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Generation of a ciliary margin-like stem cell niche from self-organizing human retinal tissue
The ciliary margin of the eye functions as a source of multipotent progenitor cells in certain organisms but whether it plays this role in humans has not been easy to study. Here the authors culture human embryonic stem cells that self-organize into retinal tissue, and show that ciliary margin-like growth zones emerge from the developing human retinal tissue and contain stem cell niches.
- Atsushi Kuwahara
- , Chikafumi Ozone
- & Yoshiki Sasai
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Quantitative high throughput screening using a primary human three-dimensional organotypic culture predicts in vivo efficacy
Tumour microenvironment affects the outcome of pharmacological anticancer treatments. Here, Kenny et al. show that organotypic cultures of ovarian cancer cells can recapitulate metastasis. They identify several new compounds that block cancer invasion and metastasis and improve survival in mouse models.
- Hilary A. Kenny
- , Madhu Lal-Nag
- & Ernst Lengyel
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Cultivation-based multiplex phenotyping of human gut microbiota allows targeted recovery of previously uncultured bacteria
Research on the roles played by gut microbes in human health is hampered by our inability to isolate and study them in pure cultures. Here, the authors describe an approach that allows targeted cultivation and rapid characterization of a significant proportion of human gut bacteria.
- Elizabeth A. Rettedal
- , Heidi Gumpert
- & Morten O.A. Sommer
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Estimating the effective density of engineered nanomaterials for in vitro dosimetry
The assessment of nanomaterial toxicity can be hampered by difficulties in determining cell dosage. Here, the authors present a simple method for determination of nanomaterial agglomerate density in liquid media, enabling accurate calculation of dose delivered to cells in an in vitrosystem.
- Glen DeLoid
- , Joel M. Cohen
- & Philip Demokritou
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Article
| Open Access
Clonal culturing of human embryonic stem cells on laminin-521/E-cadherin matrix in defined and xeno-free environment
Laminin-511 and its cell-binding domain support long-term self-renewal of human pluripotent stem cells in xeno-free culture conditions. Here the authors show that a xeno-free culture matrix consisting of a full-length laminin-521 and the cell–cell adhesion protein E-cadherin supports clonal expansion of human embryonic stem cells.
- Sergey Rodin
- , Liselotte Antonsson
- & Karl Tryggvason
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Article
| Open Access
A thermoresponsive and chemically defined hydrogel for long-term culture of human embryonic stem cells
To transfer cultured human embryonic stem cells (hESCs) between culture dishes, cells need to be released using mechanical, enzymatic or chemical means, which can damage cells. Zhanget al. describe a thermomodulatable hydrogel that allows gentle, reagent-free cell passaging for the long-term culture of hESCs.
- Rong Zhang
- , Heidi K. Mjoseng
- & Mark Bradley
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Article
| Open Access
Laminin E8 fragments support efficient adhesion and expansion of dissociated human pluripotent stem cells
The use of animal products as culture substrates for human embryonic stem cell and induced pluripotent stem cell culture raises numerous safety concerns in a therapeutic setting. Miyazaki et al.. show that minimal fragments of human laminins provide a more effective support for the culture of these cell types.
- Takamichi Miyazaki
- , Sugiko Futaki
- & Eihachiro Kawase
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Competition–colonization dynamics in experimental bacterial metacommunities
Species coexistence can be explained by the competition–colonization trade-off theory. Here, Livingston et al. illustrate this theory in a metacommunity experiment using two bacterial strains, finding a negative correlation between diversity and productivity when scaled to full metacommunities.
- George Livingston
- , Miguel Matias
- & Nicolas Mouquet
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An optimized small molecule inhibitor cocktail supports long-term maintenance of human embryonic stem cells
Stem cell-mediated regenerative medicine requires the development of defined culture systems for the maintenance of human embryonic stem cells. Here, feedback system control is used to identify a combination of three small molecule inhibitors that enables long-term human embryonic stem cell maintenance.
- Hideaki Tsutsui
- , Bahram Valamehr
- & Hong Wu
Hepatocytes differentiate into intestinal epithelial cells through a hybrid epithelial/mesenchymal cell state in culture