An optimized dissociation protocol for FACS-based isolation of rare cell types from Caenorhabditis elegans L1 larvae

Euclides E Fernandes Póvoa; Annabel L P Ebbing; Marco C Betist; Christa van der Veen; Hendrik C Korswagen

doi:10.1016/j.mex.2020.100922

An optimized dissociation protocol for FACS-based isolation of rare cell types from Caenorhabditis elegans L1 larvae

MethodsX. 2020 May 16:7:100922. doi: 10.1016/j.mex.2020.100922. eCollection 2020.

Authors

Euclides E Fernandes Póvoa¹, Annabel L P Ebbing¹, Marco C Betist¹, Christa van der Veen¹, Hendrik C Korswagen^{1

2}

Affiliations

¹ Hubrecht Institute, Royal Netherlands of Arts and Sciences and University Medical Center Utrecht, Uppsalalaan 8, 3584 CT Utrecht, Netherlands.
² Institute of Biodynamics and Biocomplexity, Developmental Biology, Department of Biology, Utrecht University, Padualaan 8, 3584 CH Utrecht, Netherlands.

Abstract

Single-cell isolation and transcriptomic analysis of a specific cell type or tissue offers the possibility of studying cell function and heterogeneity in time-dependent processes with remarkable resolution. The reduced tissue complexity and highly stereotyped development of Caenorhabditis elegans, combined with an extensive genetic toolbox and the ease of growing large tightly synchronized populations makes it an exceptional model organism for the application of such approaches. However, the difficulty to dissociate and isolate single cells from larval stages has been a major constraint to this kind of studies. Here, we describe an improved protocol for dissociation and preparation of single cell suspensions from developmentally synchronized populations of C. elegans L1 larvae. Our protocol has been empirically optimized to allow efficient FACS-based purification of large number of single cells from rare cell types, for subsequent extraction and sequencing of their mRNA.

Keywords: C. elegans; L1 larvae dissociation; Single cell isolation.