Serum-free HEK293 Cell Culture Media

Name: Serum-free HEK293 Cell Culture Media
Brand: R&D Systems
SKU: CCM023

Catalog #: CCM023 Datasheet / COA / SDS

For the serum-free expansion of human embryonic kidney (HEK) 293 cells

Discontinued Product

CCM023 has been discontinued.

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Product Details

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Summary Product Datasheets

Serum-free HEK293 Cell Culture Media Summary

Kit Summary

Media for the serum-free expansion of HEK293 cells

Key Benefits

Expands HEK293 in suspension culture
Enhances yield of expressed proteins and antibodies
Provides serum-free and low protein culture conditions
Supports a wide variety of HEK293 Cells
High lot-to-lot consistency

CCM023 Data Examples
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Why use serum-free media for the transfection of HEK293 cells?

Human Embryonic Kidney (HEK)293 cells are a commonly used mammalian cell line for the expression of recombinant proteins and antibodies. Transfecting and expanding HEK293 cells under serum-free conditions allows for the researcher to have a more controlled and defined system.

Serum-Free HEK Cell Culture Media has been optimized for the growth and expansion of HEK293 cells under serum-free, low protein (<75 µg/mL) conditions in suspension culture. This is a complete media that will support growth of a wide variety of HEK293 cells without further supplementation.

Kit Contents

This kit contains the following reagents to expand HEK293 cells in suspension culture:

Serum-free HEK Cell Culture Media (500 mL)

Stability and Storage

This media should be stored at 2 – 8 ^°C. Protect from light. Use within the expiration date listed on the bottle. For optimal culture performance, bottles should be used within 1 month after opening to prevent loss of CO₂ and elevated pH.

Precaution

This product contains human transferrin. This transferrin was tested at the donor level using an FDA licensed method and found to be non-reactive for anti-HIV-1/2 and Hepatitis B surface antigen. As no testing can offer complete assurance of freedom from infectious agents, these reagents should be handled as if capable of transmitting infection. When handling biohazardous materials such as human cells, safe laboratory procedures should be followed and protective clothing should be worn.

Limitations

FOR LABORATORY RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES.
The safety and efficacy of this product in diagnostic or other clinical uses has not been established.
This reagent should not be used beyond the expiration date indicated on the label.

Data Examples

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Serum-free HEK Cell Culture Media Supports Cell Growth of HEK293EBNA Cells. 25x10⁶ HEK293EBNA cells were inoculated in 50 mL of Serum-free HEK293 Cell Culture media. Cells were cultured at 37^°C/5% CO₂ for 7 days. The cell growth curves show the viable cell count / mL (A) and % viability (B) at each day of culture.

Specifications

Storage

Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Species

Human

Product Datasheets

Product Datasheet

COA

SDS

Assay Procedure

Refer to the product datasheet for complete product details.

Briefly, HEK293 cells are expanded using Serum-free HEK Cell Culture Media using the following procedure:

Inoculate Serum-free HEK Cell Culture Media with HEK293 cells
Incubate the shaker flask on a rotary shaker
When density of reaches 1.5 – 2.5 x 10⁶ cells/mL, expand to a larger media volume
Repeat until reaching desired expansion volume

View Graphic Procedure

Reagents Provided

Reagents provided with Serum-free HEK Cell Culture Media (Catalog # CCM023):

Serum-free HEK Cell Culture Media

Other Supplies Required

Reagents and Materials

HEK293 cells, such as HEK293EBNA
Non-baffled shake flasks
15 mL centrifuge tubes
Serological pipettes
Penicillin-Streptomycin (100X), optional

Equipment

37 °C and 5% CO₂ humidified incubator
Rotary shaker, circular orbit, 1” orbital diameter
Water bath
Centrifuge (low speed clinical or equivalent)
Hemocytometer
Microscope

Reagent Preparation:

Serum-free HEK Cell Culture Media – Serum-free HEK Cell Culture Media is complete and ready for use. If desired, add Penicillin-Streptomycin at a 1:100 dilution.

Note: If Penicillin-Streptomycin is not needed for the experiment, it can be omitted.

Protocol Overview

The protocol below describes the expansion of HEK293 cells in Serum-free HEK Cell Culture Media. Please refer to the product datasheet for complete protocol details.

Note: This procedure describes the expansion of HEK293 cells in suspension culture. Optimal seeding and passaging densities will need to be modified if using adherent cultures. Serum-free HEK293 Cell Culture Media is not designed for transfection use. For transfection it is recommended to use Serum-free HEK293 Transfection Media, Catalog # CCM024.

Warm Serum-free HEK Cell Culture Media
to 37 ^°C.

Determine the size and number of flasks needed for plating. Cells should be inoculated at 0.5 x 10⁶ cells / mL (i.e., 12.5 x 10⁶ cells in 25 mL of media in a 125mL non-baffled shaker flask).

Incubate the shaker flask on the rotary shaker at 110 – 120 RPM at 37^°C / 5% CO₂.

Count cells daily to monitor cell density and viability.

Expand cells by transferring them into a 1L non-baffled shake flask and adding Serum-free HEK Cell Culture Media to reach a final density of ~ 0.5 x 10⁶ cells / mL.

Count cells daily.

Expand cells by transferring them into a 2L non-baffled shake flask and adding Serum-free HEK Cell Culture Media to reach a final density of ~ 0.5 x 10⁶ cells / mL.

When the final desired volume is reached, continue to culture cells 3-6 days depending on application need.

Note: Final cell density at maturation is expected to reach ~ 4.0 +/- 0.5 x 10⁶ viable cells / mL by day 5-6.