Nanoparticle technology improves in-vitro attachment of cattle ( Bos taurus ) trophectoderm cells
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ORIGINAL RESEARCH PAPER
Nanoparticle technology improves in-vitro attachment of cattle (Bos taurus) trophectoderm cells Jaewook Chung . Ganesh Sriram . Carol L. Keefer
Received: 23 October 2019 / Accepted: 26 May 2020 Ó Springer Nature B.V. 2020
Abstract The bovine cell line, cow trophectoderm-1 (CT-1), provides an excellent in-vitro cell culture model to study early embryonic development. Obtaining consistent attachment and outgrowth, however, is difficult because enzymatic disassociation into single cells is detrimental; therefore, CT-1 cells must be passaged in clumps, which do not attach readily to the surface of the dish. We tested whether magnetic nanoparticles, NanoShuttleTM-PL, could be used to improve cell attachment and subsequent proliferation of the cattle trophectoderm cell line without altering cellular metabolism or immunofluorescent detection of the lineage marker Caudal Type Homeobox 2 (CDX2). Confluency was achieved more consistently by using the NanoShuttleTM-PL system to magnetically force attachment (75–100% of wells) as compared to the control (11%). Moreover, there were no alterations in characteristic morphology, nuclearlocalized expression of the trophectoderm marker CDX2, or glycolytic metabolism. By enhancing attachment, magnetic nanoparticles improved culture efficiency and reproducibility in an anchorageJ. Chung C. L. Keefer (&) Department of Animal and Avian Sciences, University of Maryland, College Park, MD 20742, USA e-mail: [email protected] G. Sriram Department of Chemical and Biomolecular Engineering, University of Maryland, College Park, MD 20742, USA
dependent cell line that otherwise was recalcitrant to efficient passaging. Keywords Trophectoderm Attachment Cell culture Magnetic nanoparticles CDX2
Introduction Cell lines derived from the trophectoderm of mammalian peri-implantation embryos have been established to aid in the study of embryonic development. One example is the CT-1 cell line, which was established from bovine embryonic trophectoderm. This line has been used extensively to study lineage differentiation and signaling between embryonic and maternal tissue (Talbot et al. 2000; Schiffmacher and Keefer 2013). Despite its value as a model for a critical embryonic stage in ruminants, the CT-1 cell line is difficult to culture due to poor attachment of passaged cell clumps. Cell clumps tend to float slightly above the surface hindering direct cell-to-surface contact, irrespective of whether that surface was formed of feeder cells or treated plastic-ware. This buoyancy is likely due to the intracellular lipid droplets which are abundant in bovine embryos (Barcelo´-Fimbres and Seidel 2007) and results in inconsistent experimental replicates as the cells do not proliferate in suspension.
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Biotechnol Lett
Therefore, one potential solution was to initiate direct surface contact by means of magnetic force. Magnetic nanoparticles have been used to levitate cells above the surface to allow formation of 3-D spheroids
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