Generation of neighbor-labeling cells to study intercellular interactions in vivo

Luigi Ombrato, Emma Nolan, Diana Passaro, Ivana Kurelac, Victoria L. Bridgeman, Alexander Waclawiczek, Delfim Duarte, Cristina Lo Celso, Dominique Bonnet, Ilaria Malanchi
Nature Protocols2021
Understanding cell–cell interactions is critical in most, if not all, research fields in biology. Nevertheless, studying intercellular crosstalk in vivo remains a relevant challenge, due mainly to the difficulty in spatially locating the surroundings of particular cells in the tissue. Cherry-niche is a powerful new method that enables cells expressing a fluorescent protein to label their surrounding cells, facilitating their specific isolation from the whole tissue as live cells. We previously applied Cherry-niche in cancer research to study the tumor microenvironment (TME) in metastasis. Here we describe how to generate cancer cells with the ability to label their neighboring cells (within the tumor niche) by transferring a liposoluble fluorescent protein. Live niche cells can be isolated and compared with cells distant from the tumor bulk, using a variety of ex vivo approaches. As previously shown, this system has the potential to identify novel components in the TME and improve our understanding of their local interactions. Importantly, Cherry-niche can also be applied to study potential cell–cell interactions due to in vivo proximity in research fields beyond cancer. This protocol takes 2–3 weeks to generate the labeling cells and 1–2 weeks to test their labeling ability.
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