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View additional product information for Leibovitz's L-15 Medium - FAQs (11415064, 11415114)
55 product FAQs found
通常情况下，加入血清后的培养基可使用三个星期。尽管没有正式的研究支持数据，这是我们研究人员的经验。
我们会在常温下运输那些需要在冰箱中长期存放的培养基。我们对代表性的培养基配方进行了研究，结果表明这些培养基在室温下放置一周不会有问题。
绝大部分情况下支原体污染无法从培养物中去除，只能弃用。不过也许您的培养物具有独特性，您不希望丢弃而试图去除污染。环丙沙星和Plasmocin据报导适合此种应用。如果对相关实验方案或应用感兴趣，请联系抗生素供应商或参考已发表的文献。请注意支原体很难从培养物中清除，而且容易扩散，所以请对受污染的培养物进行隔离处理，直至支原体被完全清除，另外您的实验室可能也需要彻底净化。
尝试更换培养基或血清。比较培养基配方中葡萄糖、氨基酸及其他成份之间的差异。比较新旧批次的血清。增加细胞的初始接种量。最后，让细胞逐步切换到新的培养基。
如果细胞经胰酶过度消化，即可能发生此种情况。尝试使用更短时间或更低浓度的胰酶进行消化处理。支原体污染也可能造成此种问题。分离部分细胞培养物，并检测支原体感染。最后，检查培养基中的粘附因子。
这最可能是细菌或真菌的污染信号。我们建议弃用培养基，同时尝试对培养物进行去污染处理。
沉淀可能是由于去垢剂漂洗过程中残留的磷酸盐所致，后者可能会沉淀带电荷的培养基成份。在去离子蒸馏水中润洗玻璃器皿数次，之后再进行灭菌。如果培养基被冰冻了，尝试将其加热至37°C后通过摇动将沉淀溶解。如果仍有沉淀，请弃用此培养基。
请参见以下pH快速波动的可能原因，及其建议的解决方案：
-不当的二氧化碳浓度：请依据培养基中的碳酸氢钠浓度来增加或降低培养箱中的二氧化碳百分比。对于2.0至3.7 g/L的碳酸氢钠，请分别使用5-10%的二氧化碳浓度。换用不依赖二氧化碳的培养基。
-组织培养瓶盖子过紧：将盖子拧1/4圈。
-碳酸氢盐缓冲不充分：加入HEPES缓冲液，使最终浓度达到10-25 mM。
-培养基中含有错误的盐分：请在CO2条件下使用Earle's盐培养基，在大气环境下使用Hanks'盐培养基。
-细菌，酵母或真菌污染：丢弃培养物及培养基。尝试对培养物进行去污染。
请参见下列常见原因，及相关问题的解决方案：
-未使用合适的生长培养基：请依照供应商推荐的培养基，预热后进行培养。
-生长培养基中的血清品质不佳：使用另一批次的血清。
-细胞传代次数过多： 请使用低传代次数的健康细胞。
-细胞铺满后的过度生长：请在长满前的对数生长期对哺乳动物细胞进行传代。
-培养物受支原体污染 ：丢弃细胞、相关培养基和试剂。获取一批新细胞，并使用新鲜培养基和试剂进行培养。
请参见下方可能原因和我们推荐的解决方案：
-细胞储存不当：获取一批新品，将其冻存于液氮中。在液氮中保存细胞直至解冻。
-自己冻存的细胞没有活性：请按照供应商推荐的密度来冻存细胞。使用低传代次数的细胞来建立冻存品。准确地遵循供应商的推荐步骤来冻存细胞。
获取一批新冻存品。
-细胞化冻操作不当：准确地遵循供应商的推荐步骤来化冻细胞。请确保快速化冻冻存细胞，但请在接种前使用预温的培养基慢慢进行稀释。
-未使用合适的化冻培养基：使用供应商推荐的培养基。确保培养基经过预热。
-细胞稀释过度：以供应商推荐的最高密度来融解细胞，以获得最佳的复苏效果。
-细胞操作不够轻柔：对大多数细胞而言，冻存和解冻的过程都是一种压力。请勿振荡或者剧烈碰撞培养瓶底来解离细胞（除非培养昆虫细胞）或高速离心细胞。
-冻存培养基中所使用的甘油在明亮环境中保存（如可行）：如果在明亮环境中保存，甘油会转化为对细胞有毒的丙烯醛。需要获取一批新品。
有许多原因都可能造成这样的后果：
不正确的CO2水平——可使用 Bacharach提供的Fyrite试剂盒(http://www.mybacharach.com/product-view/fyrite-classic/)来人工检测CO2水平,检查培养箱上显示的读数是否与人工测定的数值一致。如果培养箱显示出监测读数，则需确定CO2水平波动的结果。检查相关设置以确保CO2水平设置于适合您所用细胞系的水平（通常处于5- 10%）。经常检查管线连接以防漏气。避免频繁开关培养箱门。
培养箱中的温度波动——请在培养箱中放置一个精密的温度计来监控培养箱温度。
两性霉素或其他预防性抗生素/抗真菌素使用浓度过高——请按照推荐浓度使用。
湿度异常——检查水盘中的水量。湿度对于实现多种细胞和培养基的正常气体交换至关重要（即在湿度不足够高的条件下，对于多数培养合适的CO2水平已经无计于事）。
培养基渗透压异常——检查完全培养基的渗透压。大多数哺乳动物细胞可耐受260至350 mOsm/kg的渗透压。加入HEPES等试剂和药物可能会对渗透压造成影响。
微生物污染——细菌和真菌污染通常易于观察；支原体污染的表征微弱，不易观察, 因此对细胞培养形态的严格监控和常规测试对于检测此类污染十分必要。
使用错误的培养基——反复检查所用培养基是否准确地匹配您的细胞类型和培养应用。举例来说，请确保您所用的无血清培养基是专为无血清的培养而设计的；请确保以适当的浓度使用合适的筛选性药物；检查所用试剂的保质期；在合适的温度下将培养基储存于暗处。
如果培养基配方中含有：
NaHCO3 (g/L) 浓度<1.5时，所需CO2浓度为4%；
NaHCO3 (g/L) 浓度处于1.5-2.2时，所需CO2浓度为5%；
NaHCO3 (g/L) 浓度处于2.2-3.4时，所需CO2浓度为7%；
NaHCO3 (g/L) >3.5时，所需CO2浓度为10%。
*有个例外情况。Gibco DMEM一般遵照Dulbecco的原始配方，含有3.7g/L碳酸氢钠。用户在CO2含量为5-10%的CO2培养箱中使用此培养基已有数十年，通常可以维持生理pH值；这也与所培养的细胞类型有关。随着细胞不断生长，pH值开始不断下降（由于乳酸的代谢性积累）。
血清浓度随所用细胞系和基础培养基的不同而变化。请点击此处(https://www.thermofisher.com/us/en/home/life-science/cell-culture/mammalian-cell-culture/classical-media/advanced-d-mem-and-mem/recommended-sera-supplementations.html)查看我们推荐用于测试细胞系的血清添加剂。
有的。我们的Gibco培养基定制专家与研发专员能够为您提供添加或精简培养基成份方面的建议。如需更为全面的服务，我们的PD-Direct生物工艺服务团队将进一步为您提供培养基开发与优化的专业服务。
我们拥有Gibco定制培养基专家团队，他们能够协助您进行产品定制，生产和运输。请邮件联系custommedia@lifetech.com，或拨打www.thermofisher.com/mediaconfigurator页面列出的当地电话，以提交您的问题。
如有网络可访问性相关的问题，请通过免费或当地电话号码联系我们的系统支持。
您可在任何时间使用Gibco培养基配置工具在线浏览为您的细胞培养产品专门准备的定制选项。同时，这一工具还能够为客户定制的培养基提供更快速的询价功能，相关价格通常在48小时的工作时间内即可获得。
否，Gibco培养基配置工具无法用于非Gibco细胞培养产品的询价操作。不过，我们在帮助客户生产自制配方的培养基方面拥有非常丰富的经验。请访问www.thermofisher.com/custommedia或联系我们的 custommedia@lifetech.com 邮箱获取更多相关信息。
Gibco培养基配置工具(https://www.thermofisher.com/us/en/home/life-science/bioproduction/custom-cell-culture-media-and-services/gibco-custom-media-configurator.html)允许您增加、降低和调整各成份的浓度。此外，您还可以在包装，质控测试，以及cGMP或非cGMP生产等广泛的选项中进行选择。
几乎所有的Gibco细胞培养基础培养基产品都可通过Gibco培养基配置工具进行定制。如需检索某一特定产品，请访问培养基配置工具页面(https://www.thermofisher.com/us/en/home/life-science/bioproduction/custom-cell-culture-media-and-services/gibco-custom-media-configurator.html)，通过Gibco目录编号来进行搜索，或通过相关描述来浏览我们目前正在提供的产品。对于缓冲液和生长因子等非培养基类Gibco产品的定制服务，请通过custommedia@lifetech.com电子邮箱联系我们。
不必，您收货之后无需再调整液体培养基的pH值。这些产品已经处于正确的pH使用区间。加入添加剂（需要时）后如有需要，您可再次检测pH值并进行调整。不过，通常这一操作并不会显著改变pH值。如果您以粉剂自制培养基，则需要调整其pH值，但能够自动调节pH值和渗透压的AGT培养基不在此列。
Gibco细胞培养基用水符合USP（美国药典）对注射用水的全部要求。
您可在我们的网站上找到一份实验方案。在主搜索框中输入“以干粉和浓缩液制备培养基（Media Preparation from Powder and Concentrates）”，按下回车键或点击搜索按钮。这一操作将引导用户至一个详细的实验方案页面。
无菌技术的作用是在环境微生物与无菌的细胞培养物之间形成一道屏障，它通过一套操作流程来降低培养物被污染源污染的可能。无菌技术的组成要素包括：无菌工作区域、良好的个人卫生、无菌试剂和培养基以及无菌操作。点击此处（https://www.thermofisher.com/us/en/home/references/gibco-cell-culture-basics/aseptic-technique/aseptic-techniques-checklist.html）阅读更多关于无菌技术方面的信息，也可浏览我们的目录。
大多数常规哺乳动物细胞系在pH7.4的条件下生长良好，不同的细胞株系之间差异极小。大多数研究人员通常使用含5–7% CO2的空气，一般而言，4–10%的CO2浓度即可满足绝大多数的细胞培养实验要求。请点击此处（https://www.thermofisher.com/us/en/home/references/gibco-cell-culture-basics/cell-culture-environment/ph-co2-levels.html）阅读更多关于缓冲条件的相关信息。
请参见此处(https://www.thermofisher.com/us/en/home/references/gibco-cell-culture-basics/cell-culture-protocols/cell-culture-useful-numbers.html)的比较表。
会。动物血清应储存于–5至–20°C。培养基储存于2至8°C；请在推荐的保质期内用完。请将完全培养基（加入添加剂）保存于2 至 8°C，推荐的存放期限为2至4周。此外，尽量减少血清和培养基暴露于光线下的时间。
We do provide osmolality information on the certificate of analysis. All lots of Leibovitz's L-15 Medium will meet the osmolality specification of 300-340 mOsm/kg.
Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.
We offer the following Leibovitz media:
- Leibovitz's L-15 Medium, powder: Cat. Nos. 41300039 (10x 1L) and 41300070 (1x 10L)
- Leibovitz's L-15 Medium, no phenol red: Cat. No 21083027 (1x 500 mL)
- Leibovitz's L-15 Medium: Cat. Nos.11415064 (1x 500 mL) and 11415114 (10x 500 mL) (for sale in N. America, Latin America, and Asia-Pacific regions; Cat. No. 11415049 (for sale in Europe, Africa, and the Middle East
- Leibovitz's L-15 Medium with GlutaMAX Supplement: Cat. No. 31415029 (1x 500 mL)
Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.
Generally speaking, media can be used for up to three weeks after supplementation with serum. There are no formal studies to support this, but it is the rule of thumb used by our scientists.
Find additional tips, troubleshooting help, and resources within our Mammalian Cell Culture Basics Support Center.
We routinely ship media that require long-term storage in the refrigerator at room temperature. We have done studies on representative media formulations to show that media can be at room temperature for up to a week without a problem.
Find additional tips, troubleshooting help, and resources within our Mammalian Cell Culture Basics Support Center.
Very often mycoplasma contamination cannot be removed from the culture so it should be discarded. You may have a unique culture that you prefer not to discard and would like to try to clean it. Ciprofloxacin and Plasmocin have reportedly been used for this application. If interested in a protocol or directions for use, check with the antibiotic supplier or published literature. Note that mycoplasma are very difficult to remove from culture and spread easily so the treated cultures should be quarantined until clear of mycoplasma, and your laboratory should be thoroughly cleaned.
Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.
Try changing the medium or serum. Compare media formulations for differences in glucose, amino acids, and other components. Compare an old lot of serum with a new lot. Increase initial cell inoculums. Lastly, adapt cells sequentially to new medium.
Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.
This can occur if cells are overly trypsinized. Trypsinize for a shorter time or use less trypsin. Mycoplasma contamination could also cause this problem. Segregate your culture and test for mycoplasma infection. Lastly, check for attachment factors in the medium.
Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.
This is most likely a sign of bacterial or fungal contamination. We would recommend discarding the media, and trying to decontaminate the culture.
Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.
The precipitate could be due to residual phosphate left over from detergent washing, which may precipitate powered medium components. Rinse glassware in deionized, distilled water several times, then sterilize. If the medium is frozen, try warming media to 37 degrees C and swirl to dissolve. If precipitate remains, discard medium.
Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.
Please see the possible causes for this rapid pH shift, and suggested solutions:
- Incorrect carbon dioxide tension: Increase or decrease percentage of carbon dioxide in the incubator based on concentration of sodium bicarbonate in the medium. For sodium bicarbonate concentrations of 2.0 to 3.7 g/L, use carbon dioxide amounts of 5-10%, respectively. Switch to carbon dioxide-independent medium.
- Overly tight caps on tissue culture flasks: Loosen caps one-quarter turn.
- Insufficient bicarbonate buffering: Add HEPES buffer to a final concentration of 10-25 mM.
- Incorrect salts in medium: Use an Earle's salts-based medium in a CO2 environment and a Hanks' salts-based medium in atmospheric conditions.
- Bacterial, yeast, or fungal contamination: Discard culture and medium. Try to decontaminate culture.
Find additional tips, troubleshooting help, and resources within our Mammalian Cell Culture Basics Support Center.
Please see some common reasons below, and solutions to the issue:
- Growth medium is not correct: Use pre-warmed growth medium as recommended by the supplier.
- Serum in the growth medium is of poor quality: Use serum from a different lot.
- Cells have been passaged too many times: Use healthy, low-passage number cells.
- Cells were allowed to grow beyond confluency: Passage mammalian cells when they are in the log-phase before they reach confluency.
- Culture is contaminated with mycoplasma: Discard cells, media, and reagents. Obtain new stocks of cells, and use them with fresh media and reagents.
Find additional tips, troubleshooting help, and resources within ourMammalian Cell Culture Basics Support Center.
Please see the possible causes and solutions we recommend below:
- Cells were stored incorrectly: Obtain a new stock and store in liquid nitrogen. Keep cells in liquid nitrogen until thawing.
- Homemade freezer stock is not viable: Freeze cells at a density recommended by the supplier. Use low-passage cells to make your own freezer stocks. Follow procedures for freezing cells exactly as recommended by the supplier. Obtain a new stock.
- Cells were thawed incorrectly: Follow procedures for thawing cells exactly as recommended by the supplier. Make sure that you thaw the frozen cells quickly but dilute them slowly using pre-warmed growth medium before plating.
- Thawing medium is not correct: Use the medium recommended by the supplier. Make sure the medium is pre-warmed.
- Cells are too dilute: Plate thawed cells at the highest density recommended by the supplier to optimize recovery.
- Cells not handled gently: Freezing and thawing procedures are stressful to most cells. Do not vortex, bang the flasks to dislodge the cells (except when culturing insect cells), or centrifuge cells at high speeds.
- Glycerol used in the freezing medium was stored in light (if applicable): If stored in light, glycerol gets converted into acrolein, which is toxic to cells. Obtain a new stock.
Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.
There are a number of events that can contribute to this:
1. Incorrect CO2 levels-monitor the level of CO2 manually with a Fyrite kit, available from Bacharach (http://www.bacharach-inc.com/fyrite_analyzers.htm). Check if the manual readings concur with the readings displayed on the incubator. If the incubator has a trace readout, check the printout for fluctuations in CO2 level. Check the settings to insure that CO2 levels are set at appropriate levels for your cell line (usually between 5 and 10%). Check line connections frequently for leaks. Avoid frequent opening and closing of incubator doors.
2. Temperature fluctuations in the incubator-Monitor the temperature of incubator with a good thermometer inside the incubator.
3. Fungizone or other preventive antibiotics/antimycotics are present at toxic concentrations-use at recommended levels.
4. Humidity is incorrect-check the water level in the water pan. Humidity is vital to appropriate gas exchange for many types of cells and media (i.e., appropriate CO2 levels are largely irrelevant for most cultures if the humidity level is not high enough).
5. Incorrect osmotic pressure in medium-check osmolality of complete medium. Most mammalian cells can tolerate an osmolality of 260 to 350 mOsm/kg. Additions of reagents such as HEPES and drugs may affect osmolality.
6. Contamination by microorganisms-bacterial and fungal contaminations are usually easily visible; symptoms of mycoplasma contamination are more subtle, and careful monitoring of culture morphology and regular testing are necessary to detect this type of contamination.
7. Inappropriate medium is being used-double-check that the medium used is appropriate for your cell type and culture application. For example, ensure that the medium being used for serum-free culture is actually designed for serum-free culture; make sure that appropriate selective drugs are used at appropriate levels; check the expiration dates for the reagents being used; and store medium at appropriate temperatures in the dark.
Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.
If the media formulation contains:
- NaHCO3 (g/L) <1.5, it needs CO2 at 4%
- NaHCO3 (g/L) 1.5-2.2, it needs CO2 at 5%
- NaHCO3 (g/L) 2.2-3.4, it needs CO2 at 7%
- NaHCO3 (g/L) >3.5, it needs CO2 at 10%
However, there are some exceptions. Gibco DMEM has always been made according to Dulbecco's original published formulation, with 3.7 g/L sodium bicarbonate. Customers have been using this medium in CO2 incubators ranging from 5-10% CO2 for decades, usually with no trouble maintaining physiological pH; this also depends on the cell type. Once cells are growing, the pH will drop (due to metabolic accumulation of lactic acid).
Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.
The serum concentration will vary with the cell line and basal medium used. Please go here to see our recommended sera supplementations for tested cell lines (https://www.thermofisher.com/us/en/home/life-science/cell-culture/mammalian-cell-culture/classical-media/advanced-d-mem-and-mem/recommended-sera-supplementations.html).
Find additional tips, troubleshooting help, and resources within our Mammalian Cell Culture Basics Support Center.
Yes. Our Gibco Custom Media Specialists and R&D staff can provide advice related to the addition or elimination of media components. For more comprehensive services, our PD-Direct Bioprocess Services team specializes in media development and optimization.
Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.
We have a dedicated team of Gibco Custom Media Specialists who can partner with you during customization, manufacturing, and delivery. Please email custommedia@lifetech.com or call the local phone number listed on www.thermofisher.com/mediaconfigurator with inquiries about your submission. For questions related to website effectiveness, please contact System Support through the toll-free or local phone number.
Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.
Using the Gibco Media Configurator, you can view specific customization options for your cell culture product, online, at any time. Also, this tool provides faster pricing for custom media, generally within 48 business hours.
Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.
No, the Gibco Media Configurator cannot be used to request pricing on non-Gibco cell culture products. However, we are very experienced at manufacturing customer-owned media formulations. Please visit www.thermofisher.com/custommedia or contact us at custommedia@lifetech.com for more information.
Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.
The Gibco Media Configurator allows you to add, remove, and adjust the concentration of components. In addition, you can select from a range of packaging, QC tests, and either cGMP or non-cGMP manufacturing.
Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.
Almost all Gibco cell culture basal media products can be customized using the Gibco Media Configurator. To check on a particular product, use the Media Configurator page (https://www.thermofisher.com/us/en/home/life-science/bioproduction/custom-cell-culture-media-and-services/gibco-custom-media-configurator.html to search by Gibco catalog number, or browse our offering by description. For the customization of non-media Gibco products, such as buffers and growth factors, contact us at custommedia@lifetech.com.
Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.
No, you do not need to adjust the pH of the liquid media when you receive it. It should be in the correct pH range for use. After you add your supplements (if needed), you can check the pH again and adjust if necessary. However, usually it does not change significantly. It would be necessary to pH media that you make up yourself from a powder, with the exception of AGT media where pH and osmolality are auto-adjusted.
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Gibco cell culture media is formulated using water meeting all USP monograph requirements for Water For Injection.
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You can find a protocol on our web site. In the main search box type “Media Preparation from Powder and Concentrates” and hit Enter or click Search. This should take you to a detailed protocol page.
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Aseptic technique, designed to provide a barrier between the microrganisms in the environment and the sterile cell culture, depends upon a set of procedures to reduce the probability of contamination from these sources. The elements of aseptic technique are an aseptic work area, good personal hygiene, sterile (or sterile filtered) reagents and media, and aseptic handling. Go here to read more (https://www.thermofisher.com/us/en/home/references/gibco-cell-culture-basics/aseptic-technique.html) about aseptic technique or view our checklist (https://www.thermofisher.com/us/en/home/references/gibco-cell-culture-basics/aseptic-technique/aseptic-techniques-checklist.html).
Most normal mammalian cell lines grow well at pH 7.4, and there is very little variability among different cell strains. While most researchers usually use 5-7% CO2 in air, 4-10% CO2 is common for most cell culture experiments. Read more about buffering conditions here (https://www.thermofisher.com/us/en/home/references/gibco-cell-culture-basics/cell-culture-environment/ph-CO2-levels.html).
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Please see the comparison chart here (https://www.thermofisher.com/us/en/home/references/gibco-cell-culture-basics/cell-culture-protocols/cell-culture-useful-numbers.html).
Yes. Store animal sera at -5 to -20 degrees C. Store media at 2 to 8 degrees C; use within recommended shelf life period. Store complete media (supplemented) at 2 to 8 degrees C, and for complete medium the recommended shelf life is 2 to 4 weeks. Additionally, minimize exposure of sera and media to light.
Please use our media formulation search tool (https://www.thermofisher.com/search/results?docTypes=MediaFormulation&persona=DocSupport&linkIn=true&query=).
Find additional tips, troubleshooting help, and resources within our Mammalian Cell Culture Basics Support Center.