Initiation of a zebrafish blastula cell line on rainbow trout stromal cells and subsequent development under feeder-free conditions into a cell line, ZEB2J

Zebrafish
Jerry G XingNiels C Bols

Abstract

A continuous cell line, ZEB2, was developed from zebrafish blastula-stage embryos expressing enhanced green fluorescent protein (GFP). Originally the rainbow trout spleen cell line, RTS34st, was used as feeders to initiate and maintain the cells through several passages. ZEB2 was then grown for 2 years without feeders in L-15 with 15% fetal bovine serum (FBS) for 120 population doublings. This new cell line, ZEB2J, was heteroploid, had detectable telomerase activity, and was adherent. After growing into monolayers, some cells continued to grow into mounds. Cultures expressed Pou-2 mRNA and contained many alkaline phosphatase and a few stage-specific embryonic antigen-1-positive cells. In dishes coated with a phospholipid polymer (2-methacryloxyloxyethyl phosphorylcholine, MPC), ZEB2J formed spherical aggregates. Aggregates attached to conventional culture plastic, and most cells that emerged from aggregates had typical epithelial-like shapes of ZEB2J, which suggests that ZEB2J had limited differentiation potential, despite expressing some stem cell properties. The fluorescence of ZEB2J allowed relationships with feeder cells to be studied. In MPC dishes, ZEB2J formed mixed spheroids with RTS34st. In adherent cocultures, RTS34st...Continue Reading

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Citations

Nov 19, 2010·In Vitro Cellular & Developmental Biology. Animal·S Richelle MonaghanLucy E J Lee
May 14, 2011·Zebrafish·Vanesa RoblesJuan Carlos Izpisua Belmonte
May 16, 2013·Zebrafish·Suma ChoorapoikayilJeroen den Hertog
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Dec 21, 2013·In Vitro Cellular & Developmental Biology. Animal·Nguyen T K VoLucy E J Lee
May 20, 2014·Comparative Biochemistry and Physiology. Part A, Molecular & Integrative Physiology·Sarah J GignacLucy E J Lee
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Mar 6, 2018·The Journal of Eukaryotic Microbiology·Marcel D O Pinheiro, Niels C Bols
Jan 29, 2021·Cytotechnology·Najla Adel SalehTânia Beatriz Creczynski-Pasa

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