A SIMPLE METHOD FOR ISOLATION OF NEOBLASTS FROM PLANARIA

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CHITSAZAN M.; GOURABI H.; JABARI ARFAEI A.; BAHARVAND HOSSEIN

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Journal: Cell Journal (Yakhteh) Year:2009 | Volume:11 | Issue:2 (42) Page(s): 134-141

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Title

A SIMPLE METHOD FOR ISOLATION OF NEOBLASTS FROM PLANARIA

Author(s)

CHITSAZAN M. | GOURABI H. | JABARI ARFAEI A. | BAHARVAND HOSSEIN | Issue Writer Certificate

Keywords

STEM CELLSQ1

FLATWORMSQ1

PLANARIANQ1

REGENERATIONQ1

Abstract

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 18mm 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 11mm nylon meshes. In order to obtain purified neoblasts in the final suspension; this suspension has been compared with a cell suspension from 30Gy 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 analyses 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).

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APA: Copy

CHITSAZAN, M., GOURABI, H., JABARI ARFAEI, A., & BAHARVAND, HOSSEIN. (2009). A SIMPLE METHOD FOR ISOLATION OF NEOBLASTS FROM PLANARIA. CELL JOURNAL (YAKHTEH), 11(2 (42)), 134-141. SID. https://sid.ir/paper/16405/en

Vancouver: Copy

CHITSAZAN M., GOURABI H., JABARI ARFAEI A., BAHARVAND HOSSEIN. A SIMPLE METHOD FOR ISOLATION OF NEOBLASTS FROM PLANARIA. CELL JOURNAL (YAKHTEH)[Internet]. 2009;11(2 (42)):134-141. Available from: https://sid.ir/paper/16405/en

IEEE: Copy

M. CHITSAZAN, H. GOURABI, A. JABARI ARFAEI, and HOSSEIN BAHARVAND, “A SIMPLE METHOD FOR ISOLATION OF NEOBLASTS FROM PLANARIA,” CELL JOURNAL (YAKHTEH), vol. 11, no. 2 (42), pp. 134–141, 2009, [Online]. Available: https://sid.ir/paper/16405/en

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