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- 详细信息
- 文献和实验
- 技术资料
- 保存条件:
详询
- 库存:
99
- 供应商:
广州市左克生物
- 规格:
1x10⁶ cells / 1.0 ml
| Cat. No. | T0544 |
| Name | Immortalized Mouse Preadipocyte Cells (ScAP-23) |
| Description |
The Immortalized Mouse Preadipocyte Cells (ScAP-23) was isolated from subcutaneous adipose tissue from C57BL/6 mice and immortalized by infection with retroviruses carrying hTERT gene. The ScAP-23 preadipocytes convert to adipocytes when treated with dexamethasone, 3-methylisobutylxanthine, insulin, and indomethacin. Expression of key adipogenic transcription factors such as PPARɣ, C/EBP family, SREBP-1c transcription factors and Krox20/Egr2 are found upregulated during ScAP-23 adipogenesis. The ScAP-23 adipocytes contain abundant lipid droplets and express transcripts for mature adipocytes markers such as SCD1, aFABP, ATGL, FAS, LDL, and GPDH. Furthermore, ScAP-23 adipocytes demonstrate insulin responsiveness and express insulin receptor-β and major insulin-responsive glucose transporter GLUT4. Thus, the cell line is recommended as a tool for adipocytes biology and adipogenesis research. |
| Organism | Mouse (M. musculus) |
| Tissue | Adipose |
| Donor History | C57BL/6 mice |
| Growth Properties | Adherent, fibroblast-like |
| Cell Type | Immortalized Cells |
| Storage Condition | Vapor phase of liquid nitrogen, or below -130°C. |
| Shipping Conditions | Ship with dry ice. |
| Product Format | Frozen |
| Intended Use | This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. |
| BioSafety | II |
| Certificate of Analysis | For batch-specific test results, refer to the applicable certificate of analysis that can be found at www.abmgood.com. |
| Growth Conditions | Use of PriCoat™ T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. PriGrow III (TM003) + 2 mM L-glutamine (G275) + 10% FBS + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO₂ |
| Unpacking and Storage Instructions |
1. Visually examine the packaging containers for signs of leakage or breakage. 2. Immediately transfer frozen cells from dry ice packaging to a temperature below -130°C, preferably in liquid nitrogen vapor phase storage, until ready for use. To ensure the highest level of viability, thaw the vial and initiate culture as soon as possible upon receipt. If continued storage is desired, the vial should only be stored below -130°C or in liquid nitrogen vapor phase. Do not store at -70°C, as it will result in loss of viability. |
| Thawing Protocol |
1. Thaw cells quickly in a 37°C water bath while agitating gently (maximum 2 minutes). The vial cap should be kept above the water level to minimize the risk of contamination. 2. Decontaminate the vial by spraying and wiping the exterior of the vial with 70% ethanol. From this point onwards, all operations should be strictly carried out inside a biological safety cabinet using aseptic conditions. 3. Transfer the cell suspension into a 15ml sterile conical tube containing 5ml of pre-warmed, complete growth media. Centrifuge cells at 125xg for 5-7 minutes. 4. Aspirate the supernatant without disturbing the cell pellet. Re-suspend the cell pellet in the recommended pre-warmed, complete growth media and dispense into a T25 culture flask. 5. Incubate the cells at the recommended conditions. |
| Subculture Protocol |
Volumes given below are for a T75 flask; proportionally increase or decrease the volume as required per culture vessel size. Subculture cells once the culture vessel is 80% confluent. 1. Aspirate the culture media, and add 2-3ml of pre-warmed 0.25% Trypsin-EDTA to the culture vessel. 2. Observe the cells under a microscope to confirm detachment (typically within 2-10 minutes). Cells that are difficult to detach can be put in 37°C, for several minutes to facilitate detachment. 3. Neutralize Trypsin-EDTA by adding an equal volume of the complete growth media into the culture vessel. 4. Transfer the culture suspension into a sterile centrifuge tube, and centrifuge at 125xg for 5 minutes. The actual centrifuge duration and speed may vary depending on the cell type. 5. Aspirate the supernatant, and re-suspend the pellet with pre-warmed fresh complete growth media. Add appropriate aliquots of the cell suspension to new culture vessels, as desired. 6. Incubate the cells at the recommended conditions. |
| Cryopreservation | Cryopreservation Medium (TM024), or complete growth media with 10% DMSO. |
| Seeding Density (cells/cm2) | 20,000 - 50,000 |
| Population Doubling Time (h) | 16 - 26 |
| Immortalization Method | Serial passaging and infection with retroviruses carrying hTERT gene |
| Expression | Mature adipocytes markers (SCD1, aFABP, ATGL, FAS, LDL, and GPDH) |
| Material Citation | If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. T0544. |
| Warranty | abm warrants that cell lines shall be viable upon initiation of culture for a period of thirty (30) days after shipment and that they shall meet the specifications on the applicable abm Material Product Information sheet, certificate of analysis, and/or catalog description. Such thirty (30) day period is referred to herein as the "Warranty Period”. |
| Disclaimer |
1. All test parameters provided in the CoA are conducted using abm's standardized culture system and The stated values may vary under the end-user's culture conditions. Please verify that the product is suitable for your studies by referencing published papers or ordering RNA (0.5 μg, Cat.# C207, $450.00) or cell lysate (100 μg, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final. 2. We recommend live cell shipments for ease of cell transfer and this option can be requested at the time of order placement. Please note that the end-user will need to evaluate the feasibility of live cell shipment by taking into account the final destination's temperature variation and its geographical location. 3. All of abm's cell biology products are for research use ONLY and NOT for therapeutic/diagnostic applications. abm is not liable for any repercussions arising from the use of its cell biology product(s) in therapeutic/diagnostic or any other non-RUO application(s). 4. abm makes no warranties or representations as to the accuracy of the information on this site. Citations from literature are provided for informational purposes only. abm does not warrant that such information has been shown to be accurate. 5. abm warrants that cell lines shall be viable upon initiation of culture for a period of thirty (30) days after shipment and that they shall meet the specifications on the applicable abm Material Product Information sheet, certificate of analysis, and/or catalog description. Such thirty (30) day period is referred to herein as the "Warranty Period".
|
| Depositor | University of Toledo |
| Application | Research Use Only. |
| Material Citation | If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. T0544 |
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文献和实验实验材料: 1. 细胞来源:8—12天龄的小鼠; 2. 清洗液:不含Ca2+ 和Mg2+ 的1×PBS,添加200000IU/L青霉素、200mg/L链霉素,pH7.4; 3. 消化液:0.3%胶原酶溶液,含4%牛血清白蛋白。用Hanks液配制; 4. DW培养液:DMEM和WAJC404培养液按1:1(V/V)混合,添加100IU/ml青霉素、50µg/ml链霉素; 5. 培养液a:DW培养液,10%胎牛血清; 6. 培养液b:DW培养液,补充胰岛素10
吸管反复吹打使组织块分散,然后通过孔径25μM(200目)的筛网过滤,收集滤液和未过滤的组织块。5、将滤液以600g离心5分钟弃上清,加入原代培养的基础培养基制成细胞悬液。6、将未滤过的组织按照上述过程再处理一次,将俩次获得的细胞悬液混匀计数,按照104个/cm2的密度接种于培养瓶里,于37度,5%CO2培养箱中培养。接种12-16小时基本贴壁。在增殖状态下的前脂肪细胞可以使用胰蛋白酶消化的方法传代,并且可以冻存和复苏。
甲腺原氨酸(T3 )200pmol/L; 8. 筛网:孔径为25μm的尼龙网筛。 实验方法: 1. 取材 ① 取用普通食物喂养的4周龄雄性大鼠,乙醚麻醉后断头处死; ② 在无菌条件下从附睾周围切取脂肪垫,放入培养皿中。尽量除去血管。然后用清洗液冲洗3次; 2. 分离细胞 ① 充分剪碎所取脂肪细胞。然后放入消化液中,37℃水浴中振荡消化40
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