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How to Ensure Batch-to-Batch Consistency in Cell Culture


Are your collagen cell culture media batches consistent?

Even minor inconsistencies can cause significant issues in the lab, leading to extra testing, delays, cost overruns or even unusable output.

In this article we discuss why batch-to-batch collagen consistency is so important for cell culture and share some good practice to help ensure it doesn’t affect your results.

What is batch-batch consistency?

In a cell culture process multiple target cells are extracted, identified and incubated under conditions that are as similar as possible. This enables the output of a fixed variable to be tested and confirmed by comparing the results of each sample, and allows the development of spare cells or higher volume material. This process only works if the conditions in which the cells were grown were identical. The incubation temperature, timing, equipment used, gases introduced, reagents, and nutrients added must be the same for each test sample, as must the growth matrix.

Successful cell culture requires a matrix that can support target cell growth by keeping conditions stable, deliver the right nutrients to the most relevant locations, and enable the cells to migrate or remain fixed as the sample requires.

As we have previously discussed, collagen is one of the most effective materials used in cell culture media and is able to provide excellent conditions in which cells can develop.

However, collagen sources do not always produce cell culture media batches that provide identical conditions; known as batch-to-batch consistency.

What happens if collagen batches are inconsistent?

If batch-to-batch consistency can’t be ensured, then the cultures can become severely compromised.

Culture matrix needs to be a stable variable that can be controlled in research so that other aspects of the process or samples can be investigated as easily as possible. Even a minor change in the ionic concentration, pH level or sustained incubation temperature can render subsequent batches useless. In addition, in cell production the laboratory needs to have confidence that the output hasn’t been affected by the culture matrix, and that all of the material developed is safe and will perform as expected.

Batch-to-batch consistency is also important for regulatory compliance. During testing external analysts will assess culture matrix sources and samples to determine consistency, and if this is difficult to demonstrate the laboratory can face a greater administrative burden. These issues can ultimately lead to cell culture failure  which has knock-on impacts for laboratory costs, throughput and reputation.

So how can you minimise the chances of collagen culture matrix inconsistency affecting your processes?

Ensuring batch-to-batch consistency

The first area to look at is the transport and storage of the collagen. Collagen culture matrix must be kept at a low temperature in order to maintain integrity, and all samples should be transported and stored together in the right environment. It is also important that collagen is never used beyond the date set by the manufacturer, and any other guidance provided by the supplier should be followed to ensure compliance. Wherever possible the medium for every cell culture in a single test or production run should come from the same collagen supply to avoid any inconsistencies between deliveries.

In addition, the correct guidance should be followed when sub-culturing cell lines. If culture matrix batches are inconsistent then subsequent sub-cultures will develop under different conditions, potentially wasting the time taken to grow both the initial line and its sub-cultures.

Going right to the source

Ultimately all of the best practices that should be followed to ensure the safety of cell culture, during transit, storage and use of collagen culture matrix, will only be effective if the original source material is actually consistent between batches. As discussed in our recent white paper, collagen from widely used commercial sources can lead to batch-to-batch inconsistencies.

Typical bovine, porcine and even rat tail collagen is sourced from animals with many genetic variations and changes in their ancestry, and this can lead to minor variations between batches. Individuals of these species from which collagen is extracted can also vary significantly in size, sex, age, condition and heritage. In addition these sources, particularly bovine, can also contain viruses and pathogens that may be passed on to humans, requiring high levels of safety and stringent testing.

Our jellyfish collagen product ensures excellent levels of batch-to-batch consistency due to the relative homogeneity of individual organisms, and a conserved evolutionary structure that exhibits minimal genetic variation over millions of years.

A high quality matrix, combined with a solid culture matrix handling protocol, can help labs be more efficient and effective in every culture process.