Purification and in vitro cultivation of archaeocytes (stem cells) of the marine sponge <Emphasis Type="Italic">Hymeniacidon perleve</Emphasis> (Demospongiae) |
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Authors: | Liming Sun Yuefan Song Yi Qu Xingju Yu Wei Zhang |
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Institution: | (1) Marine Bioproducts Engineering Group,Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, China;(2) Graduate School, Chinese Academy of Sciences, Beijing, 100039, China;(3) Department of Medical Biotechnology, School of Medicine, Flinders University, Adelaide, SA 5042, Australia |
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Abstract: | Marine sponges (Porifera) are the best source of marine bioactive metabolites for drug discovery and development, although
the sustainable production of most sponge-derived metabolites remains a difficult task. In vitro cultivation of sponge cells
in bioreactors has been proposed as a promising technology. However, no continuous cell line has as yet been developed. Archaeocytes
are considered to be toti/multipotent stem cells in sponges and, when purified, may allow the development of continuous sponge
cell lines. As a prerequisite, we have developed a novel four-step protocol for the purification of archaeocytes from a marine
sponge, Hymeniacidon perleve: (1) differential centrifugation to separate large sponge cells including archaeocytes; (2) selective agglomeration in low-Ca2+/Mg2+ artificial seawater in which living archaeocytes form small loose aggregates with some pinacocytes and collencytes; (3) differential
adherence to remove anchorage-dependent pinacocytes, collencytes and other mesohyl cells; (4) Ficoll-Vrografin density gradient
centrifugation to purify archaeocytes. The final purity of archaeocytes is greater than 80%. The proliferation potential of
the archaeocytes has been demonstrated by high levels of BrdU incorporation, PCNA expression and telomerase activity. In 4-day
primary cultures, the purified archaeocytes show a 2.5-fold increase in total cell number. This study opens an important avenue
towards developing sponge cell cultures for the commercial exploitation of sponge-derived drugs.
The authors are grateful for the financial support of the Chinese Academy of Sciences under the “100 Talent Project”, the
“Innovation Fund” from the Dalian Institute of Chemical Physics, the “Hi-Tech Research and Development Program of China” (2001AA620404),
and the European Commission (project: Silicon Biotechnology). |
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Keywords: | Archaeocyte Invertebrate cell culture Marine sponge Porifera Hymeniacidon perleve (Demospongiae) |
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