Journey Into the Diverse World of Cells
Cells are demanding. Some like to be free while others like to stick around. If you're struggling with setting up and maintaining your cell cultures, you're not alone. Depending on the type of cells you're working with, different challenges can arise.
That's why we're here to help you out with some tips and tricks to overcome these challenges and keep your cells healthy and happy.
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What are the different cell types?
Examples include blood cells and some cancer cells. Culturing suspension cells can be tricky because they need specialized culture vessels and techniques to maintain their viability and growth.
One big challenge with suspension cells is aggregation, which means your cells clump together and make it difficult to maintain single-cell suspensions. To prevent this, it's important to create single-cell suspensions by gently pipetting or using specialized culture vessels.
The right culture vessel is crucial here, and different cell types may require different surfaces, such as plastic, glass, or coated surfaces.
Another challenge is confluency. Adherent cells can quickly reach confluency, meaning the entire surface of the culture vessel is covered with cells, which can lead to nutrient depletion and cell death. To avoid this, seed your cells at an appropriate density and aim for 70 to 80% confluency. Lastly, detaching adherent cells from the surface for passaging or analysis can be tough and can lead to cell damage. Try using detachment agents such as trypsin or EDTA to make this process easier.
Feeder cells can provide essential growth factors, nutrients, and signaling molecules to support the growth of cells that are difficult to culture on their own.
How can the right lab equipment and supplies help?
The right lab equipment and supplies can make all the difference to the success of your cell culture. For suspension cells, use conical tubes or specialized culture vessels to keep cells in suspension and prevent aggregation. For adherent cells, use tissue culture flasks or tissue culture dishes with the right coating to promote cell attachment and growth. And for feeder cells, use flasks or dishes with a proper surface and growth media to support their growth and function.
I need to produce monoclonal antibodies (mAbs). What vessel should I use?Bioreactor flasks
Bioreactor flasks are specialized vessels used to culture cells at a large scale, making them a valuable tool for the production of mAbs. You may already know that to produce mAbs, large quantities of hybridoma cells are needed. While hybridoma cells can be cultured in standard culture vessels, bioreactor flasks provide several advantages when scaling up the production process. Bioreactor flasks have a large surface area and volume, allowing for increased cell density and greater antibody production. They also can control environmental conditions, such as temperature, pH, and oxygen levels, to optimize cell growth and antibody production.
Frequently Asked Questions
What are the challenges of culturing primary cells compared to established cell lines?
Culturing primary cells presents unique challenges compared to established cell lines. Primary cells have a limited lifespan and demand careful handling and specialized conditions to maintain their original properties. They are usually much more sensitive to culture conditions. Customized media formulations and shorter passaging intervals are often necessary to maintain viability.
Can I culture multiple cell types together in the same flask/plate?
Yes, you can culture multiple cell types together in the same flask or plate, a technique known as co-culture. However, there are a few considerations:
● Compatibility: Ensure the cell types are compatible and won't inhibit each other's growth or function.
● Media: Develop a culture medium that supports the growth of both cell types adequately. You might need to adjust the composition to meet the nutritional requirements of each cell type. Alternatively, you may use different media for each cell type if you’re using a dedicated co-culture system such as a permeable insert.
● Cell Ratios: Determine the appropriate ratio of each cell type based on your goals and the desired level of interaction.
● Controls: Include appropriate controls, such as monocultures of each cell type, to assess the effects of co-culture.
How can I prevent cross-contamination of cells?
Ensure that vials with different cell lines are not present in the hood simultaneously, and always clean between each cell type. If feasible, use separate equipment (such as bottles and media) for each cell line, and clearly label them with the cell name. Regularly authenticate your cells using techniques like PCR. Keep accurate records of passages, freezing, and thawing to ensure cell line identity and traceability.
My cells won’t grow. What has gone wrong?
There could be many reasons why cells may not be growing. Some possible causes include:
- Incorrect seeding density
- Lack of nutrients
- Suboptimal culture conditions
- Contamination with bacteria or fungi
- Cell death due to toxic compounds
- Genetic instability
Check your protocol, including the media, growth factors, and incubation conditions, to ensure they are appropriate for the cell type being cultured. Additionally, checking for signs of contamination and performing regular cell viability assays can help identify potential issues.
Can I grow my adherent cell line in a non-treated flask?
While your cells may grow in a non-treated flask, it’s likely many cellular characteristics will change as the cells struggle to anchor to the surface. Non-treated flasks lack the coating or surface treatment that provides a hydrophilic and negatively charged environment for cell adhesion, which is necessary for the attachment and growth of adherent cells. Where possible, we would always recommend using treated flasks for anchorage-dependent cells.