Cat. №: С730п
Package: 500 ml
1 744,80 ₽
0,00 ₽
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The serum-free, protein-free nutrient medium HybriS-1-293 is intended for highly effective suspension cultivation of cells of recombinant clones and lines of НЕК-293 in biotechnological...
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Serum-free medium HybriS-1-293

The medium HybriS-1-293 is intended for highly effective suspension cultivation of cells of recombinant clones and lines of НЕК-293 in biotechnological processes.

Serum-free, protein-free nutrient medium HybriS-1-293 (for suspension culture of НЕК-293 cells) is a mixture of inorganic salts, amino acids, vitamins, glucose and synthetic serum substitutes dissolved in deionized water, sterilized by filtration through filters with pore size of 0.22 microns. The medium has chemically defined composition and does not contain products of animal or plant origin. 

Without Antibiotics.

This product is intended for research purposes.

  • Appearance: clear, yellow solution, without opalescence and sediment. 
  • pH: 7.0 to 7.4.
  • Osmolarity: 290+20 mosmol/kg.
  • Buffer capacity: at least 4.5 ml.

CULTIVATION METHOD:

  1. Open the bottle under aseptic conditions.
  2. Pour the prepared medium into culture flask or bioreactor cultivation systems.
  3. Seed cells with a density of 3-6 x 10⁵ cell/ml.

Note: in order to achieve high densities of suspension cultures of HEK-293 cells you should add the special feeding supplements. The composition of supplements is determined in accordance with the needs of the client.

Growth indicators of HEK-293T culture, in HybriS-1-293 media in the conditions of model bioreactor cultivation (by PanEco)

Laboratory equipment: shaker for CO₂-incubator Celltron (Infors, Switzerland); Erlenmeyer flasks with volume of 100–125 ml (working volume of the medium is 25-30 ml) or bioreactor vials of PanEco LLC with volume of 75 ml (working volume of the medium is 10-12 ml); mixing speed 115 - 130 rpm; 37 ⁰C; 5-6% CO₂.

Seed culture: HEK-239T (PanEco Ltd.), grown to a density of 4-5 x 10⁶ cell/ml in a stirred culture (flask, bioreactor bottle). The initial density of the culture in the passage is 2.5-6 x 10⁵ cells/ml.

Evaluation of test results: the cell culture effectiveness was evaluated starting from 72 hours after seeding. To count living cells, suspension of cells was stained with solution of trypan blue.

The maximum density of living cells in a stationary culture without supplements (batch culture) in the medium HybrisS-1-293 occurred on 4-6 days from the start of cultivation, it depending on the sowing density.

For HEK-293 T line with a seed density of 4 x 10⁵ cell/ml and with addition of L-alanyl-L-glutamine in the final concentration of 1600 mg/l, the maximum density was observed on day 5 and was 4.7–6.3 x 10⁶ cell/ml (n=10) (Fig. 1A). The average doubling time is 32-38 hours. At maximum density, the cells almost did not form aggregates.

HOW SUPPLIED: in sterile plastic wide neck bottle of 500 ml.

SHELF LIFE: 1 year.

STORAGE CONDITIONS: +2 to +8 °C in a place protected from light.

RECOMMENDED USE:

  • Do not use the product after its expiry date.
  • Use gloves, mask and clothes adapted to the manipulation of the product to avoid contamination.
  • Manipulate the product in aseptic conditions. 
  • Supplements such as antibiotics or L-Glutamine should be added as sterile supplements to the ready medium.
  • Medium should be clear and free of opalescence and sediment. 
  • Package: 500 ml
  • Sterility +
  • Glutamine: Without Glutamine
  • Documents: For research purposes
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