繁茂膜海绵原细胞的鉴别分离纯化和体外培养研究化学工程专业论文.docxVIP

AbstractAbstract Abstract Abstract Liming Sun(Biochemical Engineering) Directed by Prof．Xingju Yh Prof．Wei Zhang The lowest multi—cellular invertebrate animals·—sponges(Porifera)possess an- exceptional diversity of bioactive metabolites for new drug development．However the quantity of extractable sponge biomass available from the ocean cannot usually meet the demand for large scale research and elinical development of these compounds．Among various technologies proposed to solve this problem，／n vitro cultivation of marine sponge cells has been considered aS a promising approach．This area of research has therefore attracted great attention，although limited SUCCESS has been achieved．witll no single permanent proliferating cell line established． Archaeocytes are indispensable cell types，which are generally considered as the toti／multipotent‘stem cells’in sponges．They have the capacity to proliferate，and differentiate into other cell types．Based on this rationale We have focused our efforts in the development ofarchaeocyte culture／n vitro． To investigate whether the proliferating cells in primmorpb cultures are indeed associated with archaeocytes，a prerequisite is to develop techniques for identifying and obtaining highly pure archaeocytes． Firstly we investigated the structural characteristics of the main cell types of marine sponge Hymeniacidon perleve under light and uansmiSsion electron microscope．A protocol was produced for identifying，counting archaeocytes under light microscope．Furthermore，We used BrdU incorporation method and PCNA antibody to prove that，嚣the toti／multipotent cells of sponge，archaeocytes have Abstractgreat Abstract great potential to proliferate． Secondly,this study developed a novel four-step protocol for the purificmion of archacocytes from a marine sponge Hymeniacidon perleve：(1)differential centrifugation；(2)selective agglomeration；(3)differential adherence；(4) Fieoll—Vrografm density gradient centrifugation．The final purity of archa