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- 爱必梦(abm)
- T0015
- 2025年07月15日
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- 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. | T0015 |
| Name | Immortalized Rat Cardiomyocytes |
| Description |
Western blot was used to quantify SV40 gene expression in immortalized cell line. |
| Organism | Rat (R. norvegicus) |
| Tissue | Heart |
| Donor History | 1-2 day old rat |
| Growth Properties |
Adherent, polygonal shaped |
| 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) + 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) | 40,000 - 50,000 |
| Population Doubling Time (h) | 48-60 |
| Immortalization Method | Serial passaging and transduction with recombinant lentiviruses carrying SV40 Large T antigen |
| Expression |
SV40 detected by WB |
| 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. T0015. |
| 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 | Columbia University |
| 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. T0015 |
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文献和实验大鼠心肌细胞比乳鼠心肌难养，用无血清培养基，呈杆状，横纹清楚，在培养基里细胞不搏动。 大鼠心肌细胞分离一般是采用二型胶原酶分离，浓度为1mg/ml（用无钙台氏液配制），同时酶液里加入牛磺酸，BSA。一般采用Langendorff灌流的方法，大鼠开胸后，迅速取出心脏，放入冰的台氏液中，找到主动脉后，挂上Langendorff装置，衡流泵灌入无钙台氏液（37度），开始心脏会搏动几下，这样把心脏中的淤血挤出（此处也有人用有钙台氏液灌流，好让心脏充分搏动
0:04 成年大鼠心肌细胞原代培养4 0:52 内容简介52 1:18 手术预备78 4:06 剥离大鼠心脏246 5:05 分离心肌细胞305 10:15 培养心肌细胞615 10:56 代表性结果：心肌细胞培养物656 11:20 结论680 成年大鼠心肌细胞原代培养 本视频来源于网络，如有异议请联系我们，我们将在5个工作日内作出处理。
后，取一把眼科弯剪沿胸骨柄左下缘向上剪开肋骨，然后在切口中间横剪胸骨。这样只要左手稍顶，乳鼠的心脏就直接跳出来。然后用眼科弯镊从心脏中部直接将心室部分剪下，放入冰浴的 D-Hank's 液中。重复以上过程。取材完毕后，撤掉取材的手术器械。注：为了保证心肌细胞的活力，取心的操作过程尽量快，另外，最好把盛的心脏的培养皿放置在冰台上或者预冷的平衡盐液中。 2.3 用第二套手术器械进行下列操作。用眼科直镊和眼科弯剪把培养皿中的心脏周边的血凝块及纤维组织剔除掉，放在另一个预先装好D-Hank's液
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