Hybridoma Suspension Domestication Introduction

In view of the uncertainties associated with serum in cell culture and its impact on antibody production, serum-free suspension culture has emerged as a new trend in hybridoma cell culture. However, changing the culture mode of hybridoma cells is not easy and can greatly affect the activity of hybridoma cells. At this point, Creative Biolabs provides you with a mature hybridoma platform to save hybridoma clones and restore them to good condition for you.

Why Hybridoma Suspension Domestication is Needed?

In general, animal serum contains many nutrients required for cell proliferation in cell cultures, such as growth factors, hormones, transfer proteins, factors for apposition and spreading on the culture medium, protease inhibitors, and others. Because serum contains most of the factors that promote cell proliferation and maintenance, it is an almost universal growth additive for the cell culture of animal, human, and insect cells, among others. Although serum undeniably has saved a lot of time and effort for scientific research, it has many limitations. The composition of animal serum is complex, and its components and content often vary according to the sex, age, physiological state, and nutritional conditions of the animal. In addition, different factories and different batches produced by the same manufacturer can greatly affect the factors contained in the serum, limiting the reproducibility of cell culture and the scale of animal cell culture. This also makes a comparative analysis between experimental data from different investigators extremely difficult. In addition, the serum also contains contaminated IgG, which makes antibody purification difficult.

All these problems make the suspension culture of serum-free hybridoma particularly important. However, a sudden change in culture mode can inevitably affect the state and activity of hybridoma cells. Therefore, a reliable hybridoma suspension domestication process is essential for hybridoma rescue.

How to Perform Hybridoma Suspension Domestication?

The serum-free medium is equally suitable for parental cell culture before fusion and hybridoma cells after fusion. At the same time, serum-free culture greatly reduces the influencing factors of the experiment and guarantees the accuracy of the experiment.

In general, hybridoma suspension domestication in apposed culture needs to be carried out slowly in four main steps:

First, gradually reduce the serum content in the walled medium. The 20% serum content should be gradually reduced to 1% depending on the cell growth status. A Gradual reduction of the serum content can prevent the cells from undergoing a stress response that affects cell activity.

Second, perform serum-free suspension culture for domestication. Under suspension culture conditions, the initial inoculation density should be increased. At the same time, the serum content is reduced to 0. If the cells are in good condition, subsequent experiments could be performed.

Third, subclone the cell lines. The most stable hybridoma cell lines adapted to serum-free culture should be selected several times to obtain the most active hybridoma cell lines.

Finally, store the screened cell lines in a library.

Creative Biolabs is delighted to share with you the key points and experiences in hybridoma suspension domestication to facilitate the smooth running of your project.

All listed services and products are For Research Use Only. Do Not use in any diagnostic or therapeutic applications.

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