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Cultured pluripotent planarian stem cells retain potency and express proteins from exogenously introduced mRNAs

Kai Lei, Sean A. McKinney, Eric J. Ross, Heng-Chi Lee, View ORCID ProfileAlejandro Sánchez Alvarado
doi: https://doi.org/10.1101/573725
Kai Lei
1Stowers Institute for Medical Research, Kansas City, MO 64110, USA
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Sean A. McKinney
1Stowers Institute for Medical Research, Kansas City, MO 64110, USA
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Eric J. Ross
1Stowers Institute for Medical Research, Kansas City, MO 64110, USA
2Howard Hughes Medical Institute, Stowers Institute for Medical Research, Kansas City, MO 64110, USA
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Heng-Chi Lee
3Department of Molecular Genetics and Cell Biology, University of Chicago, Chicago, IL 60637, USA
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Alejandro Sánchez Alvarado
1Stowers Institute for Medical Research, Kansas City, MO 64110, USA
2Howard Hughes Medical Institute, Stowers Institute for Medical Research, Kansas City, MO 64110, USA
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  • ORCID record for Alejandro Sánchez Alvarado
  • For correspondence: asa@stowers.org
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Abstract

Planarians possess naturally occurring pluripotent adult somatic stem cells (neoblasts) required for homeostasis and whole–body regeneration. However, methods for culturing neoblasts are currently unavailable, hindering both mechanistic studies of potency and the development of transgenic tools. We report the first robust methodologies for culturing and delivering exogenous mRNA into neoblasts. We identified culture media for maintaining neoblasts in vitro, and showed via transplantation that the cultured stem cells retained pluripotency. By modifying standard flow cytometry methods, we developed a new procedure that significantly improved yield and purity of neoblasts. These methods facilitated the successful introduction and expression of exogenous mRNAs in neoblasts, overcoming a key hurdle impeding the application of transgenics in planarians. The tissue culture advances reported here create new opportunities to advance detailed mechanistic studies of adult stem cell pluripotency in planarians, and provide a systematic methodological framework to develop cell culture techniques for other emerging research organisms.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted March 12, 2019.
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Cultured pluripotent planarian stem cells retain potency and express proteins from exogenously introduced mRNAs
Kai Lei, Sean A. McKinney, Eric J. Ross, Heng-Chi Lee, Alejandro Sánchez Alvarado
bioRxiv 573725; doi: https://doi.org/10.1101/573725
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Cultured pluripotent planarian stem cells retain potency and express proteins from exogenously introduced mRNAs
Kai Lei, Sean A. McKinney, Eric J. Ross, Heng-Chi Lee, Alejandro Sánchez Alvarado
bioRxiv 573725; doi: https://doi.org/10.1101/573725

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