Preparation of single-cell suspensions from the human placenta

Valeria Garcia-Flores, Yi Xu, Errile Pusod, Roberto Romero, Roger Pique-Regi, Nardhy Gomez-Lopez

Published: 2022-11-29 DOI: 10.1038/s41596-022-00772-w

single-cell suspensions

protocol

cryopreservation

human placenta

scRNA-seq

AI 解读

Genetic manipulation and immortalized culture of ex vivo primary human germinal center B cells

The cell-line-derived subcutaneous tumor model in preclinical cancer research

Subcellular patch-clamp techniques for single-bouton stimulation and simultaneous pre- and postsynaptic recording at cortical synapses

NovoSpaRc: flexible spatial reconstruction of single-cell gene expression with optimal transport

Genome-wide pooled CRISPR screening in neurospheres

High-throughput cultivation and identification of bacteria from the plant root microbiota

Viral crosslinking and solid-phase purification enables discovery of ribonucleoprotein complexes on incoming RNA virus genomes

Mitochondrial single-cell ATAC-seq for high-throughput multi-omic detection of mitochondrial genotypes and chromatin accessibility

Addendum: Micro-CT acquisition and image processing to track and characterize pulmonary nodules in mice

Multiplexed single-cell proteomics using SCoPE2

查看全部

Sections

Figures

References

Single-cell RNA-sequencing (scRNA-seq) allows the characterization of cellular composition and interactions in complex tissues. An essential prerequisite for scRNA-seq is the preparation of high-quality single-cell suspensions. So far, no protocols have b
Introduction
Materials
Procedure
Troubleshooting
- Genotyping for demultiplexing single-cell libraries
- scRNA-seq data analysis
Timing
Anticipated results
- References
References
ACKNOWLEDGMENTS
ACKNOWLEDGMENTS

Schroeder, A. et al. The RIN: an RNA integrity number for assigning integrity values to RNA measurements. BMC Mol. Biol. 7, 3 (2006).
Aran, D. et al. Reference-based analysis of lung single-cell sequencing reveals a transitional profibrotic macrophage. Nat. Immunol. 20, 163–172 (2019).
Stuart, T. et al. Comprehensive integration of single-cell data. Cell 177, 1888–1902.e1821 (2019).
Gomez-Lopez, N. et al. Specific inflammatory microenvironments in the zones of the fetal membranes at term delivery. Am. J. Obstet. Gynecol. 205, 235.e215–224 (2011).
Burton, G. J. & Jauniaux, E. What is the placenta. Am. J. Obstet. Gynecol. 213, S6 e1 S6–S8 (2015).
Ander, S. E., Diamond, M. S. & Coyne, C. B. Immune responses at the maternal–fetal interface. Sci. Immunol. 4, eaat6114 (2019).
Redline, R. W. The clinical implications of placental diagnoses. Semin. Perinatol. 39, 2–8 (2015).
Motomura, K. et al. RNA sequencing reveals distinct immune responses in the chorioamniotic membranes of women with preterm labor and microbial or sterile intra-amniotic inflammation. Infect. Immun. 89, e00819–e00820 (2021).
Kulkarni, A., Anderson, A. G., Merullo, D. P. & Konopka, G. Beyond bulk: a review of single cell transcriptomics methodologies and applications. Curr. Opin. Biotechnol. 58, 129–136 (2019).
Haque, A., Engel, J., Teichmann, S. A. & Lonnberg, T. A practical guide to single-cell RNA-sequencing for biomedical research and clinical applications. Genome Med. 9, 75 (2017).
Pique-Regi, R. et al. Single cell transcriptional signatures of the human placenta in term and preterm parturition. eLife 8, e52004 (2019).
Pique-Regi, R. et al. Does the human placenta express the canonical cell entry mediators for SARS-CoV-2. eLife 9, e58716 (2020).
Garcia-Flores, V. et al. Maternal–fetal immune responses in pregnant women infected with SARS-CoV-2. Nat. Commun. 13, 320 (2022).
Xu, Y., Plazyo, O., Romero, R., Hassan, S. S. & Gomez-Lopez, N. Isolation of leukocytes from the human maternal–fetal interface. J. Vis. Exp. https://doi.org/10.3791/52863 (2015).
Tsang, J. C. H. et al. Integrative single-cell and cell-free plasma RNA transcriptomics elucidates placental cellular dynamics. Proc. Natl Acad. Sci. USA 114, E7786–E7795 (2017).
Pavličev, M. et al. Single-cell transcriptomics of the human placenta: inferring the cell communication network of the maternal–fetal interface. Genome Res. 27, 349–361 (2017).
Vento-Tormo, R. et al. Single-cell reconstruction of the early maternal–fetal interface in humans. Nature 563, 347–353 (2018).
Liu, Y. et al. Single-cell RNA-seq reveals the diversity of trophoblast subtypes and patterns of differentiation in the human placenta. Cell Res. 28, 819–832 (2018).
Huang, J. et al. Single-cell transcriptomics analysis showing functional heterogeneity in decidual stromal cells during labor. J. Investig. Med. 69, 851–856 (2020).
Du, L. et al. Single-cell transcriptome analysis reveals defective decidua stromal niche attributes to recurrent spontaneous abortion. Cell Prolif. 54, e13125 (2021).
Wang, F. et al. Single-cell immune landscape of human recurrent miscarriage. Genomics Proteom. Bioinforma. 19, 208–222 (2021).
Chen, P. et al. The immune atlas of human deciduas with unexplained recurrent pregnancy loss. Front. Immunol. 12, 689019 (2021).
Zhang, T. et al. Dissecting human trophoblast cell transcriptional heterogeneity in preeclampsia using single-cell RNA sequencing. Mol. Genet. Genom. Med. 9, e1730 (2021).
Zeb, Q., Wang, C., Shafiq, S. & Liu, L. in Single-Cell Omics (eds. Barh, D. & Azevedo, V.) 101–135 (Academic Press, 2019).
10x Genomics. What is the range of compatible cell sizes? https://kb.10xgenomics.com/hc/en-us/articles/218170543-What-is-the-range-of-compatible-cell-sizes- (2022).
Pique-Regi, R. et al. A single-cell atlas of the myometrium in human parturition. JCI Insight 7, e153921 (2022).

Preparation of single-cell suspensions from the human placenta

Extended

Supplementary information

推荐阅读