%0 Journal Article %T A Simple Method For Isolation Of Neoblasts From Planaria %J Cell Journal (Yakhteh) %I Royan Institute, Iranian Academic Center for Education Culture and Research (ACECR) %Z 2228-5806 %A Chitsazan, Hamed %A Gourabi, Hamid %A Arfae, Ali Jabbary %A Baharvand, Hossein %D 2009 %\ 07/01/2009 %V 11 %N 2 %P 134-141 %! A Simple Method For Isolation Of Neoblasts From Planaria %K Stem cells %K Flatworms %K Planarian %K regeneration %R %X Objective: Freshwater planarians were used as models for studying metazoan regeneration and stem cell biology. Here a simple, fast and high throughput method for extracting their stem cells (neoblasts) is represented. Materials and Methods: Specimens of the Dugesia sp with an average length of 18 mm were homogenized by a glass Dounce tissue grinder which contained about 1 ml of planarian saline solution. The extracted suspension was serially filtered by 60, 41, 30, 20 and 11 μm nylon meshes. In order to obtain purified neoblasts in the final suspension; this suspension has been compared with a cell suspension from 30 Gy irradiated worms. Hoechst 33342 was used to determine cells from non-cellular particles; methylene blue and propidium iodide were used to detect the number of dead cells in each extraction. Results: About 2.6-3 million cells were extracted from 10-12 worms. Flow cytometry analysis showed about 83% of the extracted particles were cells. In suspensions from irradiated animals, about 50% of the cells were absent, the final suspension contained about 62-66% neoblasts and about 17% non-cellular particles. When these extracts were treated with distilled water to destroy the cells, only rabdites and chitinous spines of the parenchyma were observed in the extract. Conclusion: Results show that the purity of neoblasts in the final suspension is about 66%. Non-cellular particles have a carbohydrate nature and, therefore, this extraction method is completely compatible with molecular (e.g. proteomics and transcriptomics) and cellular methods (e.g. neoblast culture). %U https://www.celljournal.org/article_248113_a304f8e80198a7a99e4e21c1f48b4b27.pdf