计算溶液所需的质量、体积或浓度。
HEK 293培养基，含谷氨酰胺
有货
为什么选择此级别
BioReagent,无菌过滤,用于细胞培养 级 ，适用于对基线干扰要求严格的色谱和分析工作流程。
储存与运输
2-8°C储存,避光。低温运输 。请查阅批次 COA 获取详细规格。
质量文档
SDS、COA、产品数据表及规格说明书均可下载。可通过批号查询获取批次 COA。
文献证明
在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
概述
本产品是化学成分限定的培养基，无血清、无蛋白或生长因子、无动物源成分、无水解物，适用于悬浮培养的HEK 293细胞，以实现细胞快速增殖、高密度培养基高效的瞬时转染。
主要成分
由水、葡萄糖、氨基酸、维生素、无机盐、微量元素、碳酸氢钠及酸碱缓冲液组成；
无血清、无蛋白 / 生长因子、无动物源成分、无水解物、无 HT、无酚红、化学成分限定；
含 4 mM 谷氨酰胺。
本产品适用于悬浮培养的 HEK293 细胞。
使用说明
1. 基础培养条件
培养温度：37.0±0.5℃（可根据细胞培养工艺调整）；CO₂浓度：5~8%；培养湿度：≥80%；摇床转速：110~150 rpm（振幅 50 mm）。
2. 细胞复苏
2.1 将本培养基置于 37℃避光条件下充分预热，取 10 mL 预热后的培养基至 50 mL 离心管备用；
2.2 取冻存细胞管于 37℃水浴中快速解冻（＜2 min）；
2.3 将解冻后的细胞液转移至备用离心管中，300 g 离心 3 min，弃上清；取 10 mL 预热培养基重悬细胞，取样计数并检测细胞密度与活率；
2.4 将细胞悬液转移至摇瓶 / 摇管中，用预热培养基稀释至所需密度，按基础培养条件进行培养。
3. 细胞传代
3.1 将本培养基置于 37℃避光条件下充分预热；
3.2 选取对数生长期，或活细胞密度≥1.5×10⁶ cells/mL 且活率≥90%的细胞，按实验需求密度进行传代；
3.3 推荐细胞接种密度：0.3~0.7×10⁶ cells/mL；
3.4 常规培养 3~5 天后，可进行下一次传代操作。
4. 细胞冻存
4.1 配制冻存液：以本培养基为基底，添加 DMSO 至终体积浓度 10%，2~8℃预冷备用；
4.2 选取对数生长期且活率≥90% 的细胞，推荐最终冻存密度：1.0~3.0×10⁷ cells/ml；
4.3 按冻存密度计算所需细胞体积，300 g 离心 5 min 收获细胞，用预冷的冻存液重悬细胞沉淀；
4.4 将细胞悬液立即分装至低温冻存管中，按标准程序冷冻（冷冻速率 - 1℃/min），在控温低温保存设备中冻至 - 80℃，并于 - 80℃冰箱过夜保存；
4.5 细胞长期储存需转移至液氮中。
5. 细胞驯化
本产品支持直接驯化与梯度驯化两种方式，驯化成功判定标准：完全使用本培养基培养 3~4 天后，细胞密度≥3.0×10⁶ cells/ml 且活率≥90%。
5.1 直接驯化：将细胞直接接种至本培养基中，接种密度参考传代密度（0.3~0.7×10⁶ cells/mL），连续传 3~6 代后细胞可稳定生长；
5.2 梯度驯化：若直接驯化效果不佳，选取低代次、对数生长期的细胞，按以下比例梯度替换培养基，每次传代接种密度保持 0.4~0.6×10⁶ cells/mL，且需至少连续 2 代达到VCD≥3×10⁶ cells/ml 且 VIA≥90%，方可进行下一步梯度替换。
| 梯度 | 本产品：原始培养基 |
| 0 | 0:100 |
| 1 | 25:75 |
| 2 | 50:50 |
| 3 | 75:25 |
| 4 | 100:0 |
6. 瞬时转染
本说明为通用转染条件，最佳转染参数需根据实验场景调整，可通过 DOE 实验优化；以下以10 mL 转染体积、细胞密度 (3.0±0.5)×10⁶ cells/mL、DNA:10 μg、DNA:PEI=1:4为例说明操作流程。
通用转染参数推荐：
| 参数 | 推荐范围 |
| 细胞密度（VCD） | 2.0~4.0×10⁶ cells/ml |
| DNA 用量 | 1.0~2.0 mg/L |
| DNA:PEI 比例 | 1：3~1：6 |
转染操作流程
6.1 用本培养基传代扩增 HEK293 细胞，至细胞密度≥(3.0±0.5)×10⁶ cells/mL 且活率≥95%；
6.2 转染前 1 天，按 1.5×10⁶ cells/ml 密度计算细胞液体积，300 g 离心 5 min，弃全部上清，用本培养基重悬细胞沉淀，按基础培养条件培养；
6.3 转染当天取样计数，确认细胞密度 (3.0±0.5)×10⁶ cells/ml、活率≥95% 后进行转染；若密度过高，用本培养基稀释至该密度（多余细胞弃用，勿用于常规传代）；
6.4 试剂稀释：用 0.45 mL 本培养基稀释 10 μg DNA，轻吹打 5~10 次混匀，室温静置 5~10 min；同时用 0.45 mL 本培养基稀释 40 μg PEI，同法处理；
6.5 复合物制备：将稀释后的 PEI 缓慢加入稀释的 DNA 中，轻吹打 5~10 次混匀，室温静置 10~15 min；
6.6 转染：将 DNA-PEI 复合物缓慢逐滴加入细胞悬液中，边加边混匀，确保复合物分散均匀；
6.7 培养：将细胞放回摇床，按基础培养条件继续培养；
6.8 后续监测：转染后第 2 天开始取样计数，检测葡萄糖浓度并维持在 4 g/L 左右；
6.9 收获：转染后 5~7 天为蛋白表达高峰期，建议培养 6~9 天或细胞活率＜60% 时，停止培养并收获样品。
注意事项
本产品出厂前已通过 0.22/0.45 μm 无菌过滤，不建议二次过滤；
产品含 4 mM 谷氨酰胺，可直接使用；也可根据 HEK293 细胞系的具体需求额外补加；
产品不含 ACA，可直接使用；也可根据 HEK293 细胞系的具体需求额外补加；
若产品在 2~8℃冰箱长期放置出现轻微浑浊，排除污染风险后，可判定为冰箱温控不均导致；将产品置于室温或 37℃水浴锅预热复溶，恢复澄清后可正常使用；
培养基使用过程中请勿反复预热，产品开封后建议 30 天内使用完毕；
细胞驯化过程中，前 3 代细胞状态可能不佳，第 4 代开始细胞状态会逐步改善。
This product is a chemically defined medium, free of serum, proteins, growth factors, animal-derived components, and hydrolysates. It is suitable for the suspension culture of HEK 293 cells to achieve rapid cell proliferation, high-density culture, and efficient transient transfection.
Main Components
Composed of water, glucose, amino acids, vitamins, inorganic salts, trace elements, sodium bicarbonate, and a pH buffer system.
Free of serum, proteins/growth factors, animal-derived components, hydrolysates, HT, and phenol red; chemically defined.
Contains 4 mM Glutamine.
This product is suitable for the culture of HEK 293 cells in suspension.
Instructions for Use
1. Basal Culture Conditions
Culture Temperature: 37.0 ± 0.5°C (can be adjusted according to the cell culture process).
CO₂ Concentration: 5~8%.
Humidity: ≥80%.
Shaker Speed: 110~150 rpm (with an amplitude of 50 mm).
2. Cell Thawing and Recovery
2.1 Pre-warm the medium completely at 37°C, protected from light. Transfer 10 mL of the pre-warmed medium into a 50 mL centrifuge tube and set aside.
2.2 Quickly thaw the frozen cell vial in a 37°C water bath (for less than 2 min).
2.3 Transfer the thawed cell suspension to the prepared centrifuge tube containing the medium. Centrifuge at 300 g for 3 min and discard the supernatant. Resuspend the cell pellet in 10 mL of pre-warmed medium. Take a sample to count cells and determine cell density and viability.
2.4 Transfer the cell suspension to a shake flask/tube. Dilute to the desired seeding density with pre-warmed medium and culture according to the basal culture conditions.
3. Cell Passaging (Subculture)
3.1 Pre-warm the medium completely at 37°C, protected from light.
3.2 Select cells in the logarithmic growth phase, or with a viable cell density (VCD) ≥ 1.5 × 10⁶ cells/mL and viability ≥ 90%. Subculture at the desired seeding density according to experimental requirements.
3.3 Recommended cell seeding density: 0.3~0.7 × 10⁶ cells/mL.
3.4 After routine culture for 3-5 days, perform the next subculture.
4. Cell Cryopreservation
4.1 Prepare Freezing Medium: Use this medium as the base and add DMSO to a final concentration of 10% (v/v). Pre-cool the mixture at 2-8°C until ready for use.
4.2 Select cells in the logarithmic growth phase with viability ≥ 90%. Recommended final cryopreservation density: 1.0~3.0 × 10⁷ cells/mL.
4.3 Calculate the required cell suspension volume based on the desired cryopreservation density. Centrifuge the required volume of cells at 300 g for 5 min to pellet them. Discard the supernatant and resuspend the cell pellet gently in the pre-cooled freezing medium.
4.4 Immediately aliquot the cell suspension into pre-chilled cryogenic vials. Freeze following a standard slow-freezing program (cooling rate of -1°C/min) using a controlled-rate freezing apparatus until reaching -80°C. Then transfer the vials to a -80°C freezer for overnight storage.
4.5 For long-term storage, cells must be transferred to liquid nitrogen.
5. Cell Adaptation (Acclimation)
This product supports both direct and gradual adaptation methods. The criteria for successful adaptation are: after 3-4 days of culture using 100% of this medium, the cell density should reach ≥ 3.0 × 10⁶ cells/mL with viability ≥ 90%.
5.1 Direct Adaptation: Inoculate cells directly into this medium at a seeding density within the recommended range for passaging (0.3~0.7 × 10⁶ cells/mL). Cells should achieve stable growth after 3-6 consecutive passages.
5.2 Gradual Adaptation: If direct adaptation is not optimal, select low-passage, logarithmically growing cells. Replace the medium gradually according to the ratios below. Maintain a seeding density of 0.4~0.6 × 10⁶ cells/mL for each passage during the adaptation process. Proceed to the next gradient only after the cells have achieved VCD ≥ 3 × 10⁶ cells/mL and VIA ≥ 90% for at least two consecutive passages.
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6. Transient Transfection
These instructions provide general transfection conditions. Optimal parameters should be determined based on the specific experimental setup and can be optimized using Design of Experiments (DOE). The following procedure uses a 10 mL transfection volume, cell density of (3.0 ± 0.5) × 10⁶ cells/mL, 10 μg DNA, and a DNA:PEI ratio of 1:4 as an example.
General Transfection Parameter Recommendations:
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Transfection Procedure
6.1 Expand HEK 293 cells using this medium until the cell density reaches ≥ (3.0 ± 0.5) × 10⁶ cells/mL with viability ≥ 95%.
6.2 Day before transfection: Calculate the required cell suspension volume based on a seeding density of 1.5 × 10⁶ cells/mL. Centrifuge the calculated volume of cells at 300 g for 5 min. Discard the entire supernatant. Resuspend the cell pellet in fresh, pre-warmed medium. Culture according to the basal conditions.
6.3 Day of transfection: Take a sample and count cells. Confirm that the cell density is (3.0 ± 0.5) × 10⁶ cells/mL and viability is ≥ 95% before proceeding with transfection. If the density is too high, dilute the cells to the target density using fresh, pre-warmed medium (discard excess cells; do not use them for routine passaging).
6.4 Reagent Dilution: Dilute 10 μg of DNA in 0.45 mL of this medium. Mix gently by pipetting 5-10 times. Incubate at room temperature for 5-10 min. Simultaneously, dilute 40 μg of PEI in 0.45 mL of this medium. Mix using the same method.
6.5 Complex Formation: Slowly add the diluted PEI solution to the diluted DNA solution. Mix gently by pipetting 5-10 times. Incubate at room temperature for 10-15 min.
6.6 Transfection: Slowly add the DNA-PEI complex dropwise to the cell suspension. Gently swirl or shake the culture vessel while adding to ensure even distribution of the complexes.
6.7 Culture: Return the culture vessel to the shaker and continue incubation under the basal culture conditions.
6.8 Monitoring: Starting from day 2 post-transfection, take samples daily for cell counting and glucose measurement. Maintain glucose concentration around 4 g/L by supplementing if necessary.
6.9 Harvesting: Protein expression typically peaks 5-7 days post-transfection. It is recommended to stop the culture and harvest the supernatant 6-9 days post-transfection, or when cell viability drops below 60%.
Notes
- This product has been sterilized by 0.22/0.45 μm filtration prior to shipping. Re-filtering is not recommended.
- The product contains 4 mM Glutamine and can be used directly. Additional supplementation may be applied based on the specific requirements of the HEK 293 cell line used.
- The product does not contain ACA and can be used directly. Additional supplementation may be applied based on specific requirements.
- If slight turbidity or precipitation appears after long-term storage at 2-8°C, after ruling out contamination, this is likely due to temperature fluctuations within the refrigerator. To remedy this, warm the bottle to room temperature or in a 37°C water bath to re-dissolve the components. The medium should return to a clear state and can be used normally.
- Avoid repeatedly warming up the medium during use. It is recommended to use the entire contents within 30 days after opening the bottle.
- During the cell adaptation process, cell performance may be suboptimal for the first 3 passages. Cell characteristics are expected to gradually improve starting from the 4th passage onwards.
规格
别名
HEK293 悬浮培养专用无血清化学成分限定培养基 | 293无血清培养液(含L-谷氨酰胺)
英文别名
293 Cell Culture Medium（Serum-free） | Serum-Free Chemically Defined HEK293 Suspension Culture Medium with 4 mM L-Glutamine
规格或纯度
BioReagent,用于细胞培养,无菌过滤
稳定性与储存
Store at 2-8℃ long term (12 months). Store in the dark.
英文名称
HEK 293 Medium, with Glutamine
储存条件
2-8°C储存,避光
运输条件
低温运输
产品线
生物产品
名称和识别符
分子类型
培养基
技术文档
高级数据
化学和物理性质
敏感性
Light-sensitive
质检证书（CoA,COO,BSE/TSE 和分析图谱)
C of A & Other Certificates(BSE/TSE, COO):
输入批号以搜索分析图谱:
通过匹配包装上的批号来查找并下载产品的 COA，每批产品都进行了严格的验证，您可放心使用！
| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 ZJ26F0535256 |
分析证书 | H1492269 |
| ZJ26F0535257 |
分析证书 | H1492269 |
技术文档和文章
用于细胞培养
目录： 阿拉丁试剂的规格、级别和纯度
108种实验室常见的细胞系
目录： 技术文章
溶液计算器
常见问题
什么是「无菌过滤」？
「无菌过滤」表示该物料经过加工和检测，适用于特种工业加工。规格侧重于影响该应用性能的杂质——例如，电子应用的金属含量、电池应用的粒度、光学应用的折射率，或对水分敏感工艺的水分含量。
什么是「BioReagent」？
「BioReagent」表示该产品经过加工和检测，适用于生物和生化应用。规格可能包括内毒素限值、微生物控制、无菌性、低 DNase/RNase/蛋白酶活性，以及在相关情况下的生物活性验证。
本产品应如何储存？
请于2–8 °C、避光条件下保存。本品对光敏感，请保存在原始的避光或棕色容器中。
本产品如何运输？
本产品采用湿冰冷藏运输。收货后请立即拆包，并按上述储存条件保存。
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每个订单都会随附分析证书（COA），您也可以从账户订单历史中下载。如需订购前的COA，请联系您的客户代表，或使用PDP上的"申请COA"按钮——我们将提供代表性批次的COA。
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