title: Purification and in vitro cultivation of archaeocytes (stem cells) of the marine sponge Hymeniacidon perleve (Demospongiae).
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-Ca(2+)/Mg(2+) 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.
authors.affiliation: Marine Bioproducts Engineering Group, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China.
authors.forename: Liming
authors.initials: L
authors.lastname: Sun
authors.forename: Yuefan
authors.initials: Y
authors.lastname: Song
authors.forename: Yi
authors.initials: Y
authors.lastname: Qu
authors.forename: Xingju
authors.initials: X
authors.lastname: Yu
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
mesh_terms: D000818:Animals; D001973:Bromodeoxyuridine; D002452:Cell Count; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D002469:Cell Separation; D011161:Porifera / Q000166:cytology* / Q000648:ultrastructure; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology* / Q000648:ultrastructure; D019098:Telomerase / Q000378:metabolism; D013997:Time Factors
chemical_list: D018809:Proliferating Cell Nuclear Antigen; D019098:Telomerase; D001973:Bromodeoxyuridine