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微信小章| 产品名称： | OP9 |
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| 商品货号： | TS209457 |
| Organism： | Mus musculus, mouse |
| Tissue： | bone marrow/stroma |
| Cell Type： | Embryonic Stem Cells, Macrophage |
| Product Format： | frozen |
| Morphology： | fibroblast-like |
| Culture Properties： | adherent |
| Biosafety Level： | 1
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. |
| Age： | embryo |
| Strain： | (C57BL/6 x C3H)F2 -op/op |
| Applications： | OP9 cells can be used to coculture mouse embryonic stem cells (ES cells) to induce the differentiation of embryonic stem (ES) cells into blood cells of erythroid, myeloid, and B cell lineages. Cocultivation with OP9 does not require exogenous growth factors or complex embryoid structures. This system will facilitate the study of molecular mechanisms involved in development and differentiation of hematopoietic cells. |
| Storage Conditions： | liquid nitrogen vapor phase |
| Images： |  |
| Derivation： | The OP9 cell line was established from newborn op/op mouse calvaria. |
| Comments： | The cells do not produce functional macrophage colony-stimulating factor (M-CSF) due to an osteopetrotic mutation in the gene encoding M-CSF. The presence of M-CSF had inhibitory effects on the differentiation of embryonic stem (ES) cells to blood cells other than macrophages.xa0 |
| Complete Growth Medium： | The base medium for this cell line is Alpha Minimum Essential Medium without ribonucleosides and deoxyribonucleosides and with 2.2 g/L sodium bicarbonate. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 20%
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| Subculturing： | Volumes are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes.
Note: Cell density is important. If the subculture ratio is too low, the culture will not reach confluence. However, do not overgrow. Very large cells tend to appear after overgrowth and these cells are a warning sign that the OP9 cells will not support the maintenance of hematopoietic cells. Subculture just before confluence.
Interval: Maintain cultures at a cell concentration between 4 X 103 and 1 X 104xa0cells/cm2.
Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:5 is recommended
Medium Renewal: Every 2 to 3 days |
| Cryopreservation： | Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase |
| Culture Conditions： | Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37°C |
| Population Doubling Time： | 26 hrs |
| Name of Depositor： | T Nakano |
| Deposited As： | Mus musculus |
| References： | Nakano T, et al. Generation of lymphohematopoietic cells from embryonic stem cells in culture. Science 265: 1098-1101, 1994. PubMed: 8066449 Nakano T, et al. In vitro development of primitive and definitive erythrocytes from different precursors. Science 272: 722-724, 1996. PubMed: 8614833 Nakano T. Lymphohematopoietic development from embryonic stem cells in vitro. Semin. Immunol. 7: 197-203, 1995. PubMed: 7579206 Motoyama N, et al. bcl-x prevents apoptotic cell death of both primitive and definitive erythrocytes at the end of maturation. J. Exp. Med. 189: 1691-1698, 1999. PubMed: 10359572 Nakano T. In vitro development of hematopoietic system from mouse embryonic stem cells: a new approach for embryonic hematopoiesis. Int. J. Hematol. 65: 1-8, 1996. PubMed: 8990620 Nakano T, et al. Development of erythroid cells from mouse embryonic stem cells in culture: potential use for erythroid transcription factor study. Leukemia 3: 496-500, 1997. PubMed: 9209437 Suwabe N, et al. GATA-1 regulates growth and differentiation of definitive erythroid lineage cells during in vitro ES cell differentiation. Blood 92: 4108-4118, 1998. PubMed: 9834216 Suzuki A, Nakano T. Development of hematopoietic cells from embryonic stem cells. Int. J. Hematol. 73: 1-5, 2001. PubMed: 11372743 Eto K, et al. Megakaryocytes derived from embryonic stem cells implicate CalDAG-GEFI in integrin signaling. Proc. Natl. Acad. Sci. USA 99: 12819-12824, 2002. PubMed: 12239348 Freshney RI. Culture of Animal Cells: A Manual of Basic Technique, 5th edition. New York: Wiley Liss; 2005. For more information on enzymatic dissociation and subculturing of cell lines see Chapter 13. |