Fragile Immune Cells Require Gentle Handling

Updated on Oct 2, 2023 Share

Protecting cells during cell processing is a critical aspect of protocol design and development. Cells are fragile by nature, requiring specific and gentle handling to survive and optimally perform in their downstream applications. Some cells are more delicate than others and more prone to damage or cell death when exposed to stressful conditions.

The Importance of Gentle Handling

Handling cells gently is crucial to maintaining cell viability, morphology, and experimental consistency. Cells are delicate structures prone to damage when disrupted. Retaining the shape and inner structure throughout processing is integral to retaining their functionality and integrity, as some cell types utilize their morphology to carry out immune functions. 

Gentle handling preserves cell viability, ensuring survival in downstream applications. Additionally, by minimizing manipulation, outside contamination and error are less likely to occur, increasing the replicability of the cell separation process. 

Many experiments require intact cell structures and distinct morphology for precise results. Preserving this morphology through gentle handling is essential for observation-based microscopic applications, histology, and various imaging techniques. Overall gentle handling preserves viability and morphology while increasing sample consistency. 

Common Fragile Cell Types

Stem Cells

Stem cells are undifferentiated cells that divide and provide a constant supply of differentiated somatic cells. Their undifferentiated nature makes them highly sensitive to changes in their microenvironment. This heightened sensitivity requires hyper-specific culture media and growth factors to maintain their self-renewal qualities, which are commonly studied in stem cell applications. 

T cells

T lymphocytes are the activators of the inflammatory immune response, distinguishing pathogenic from commensal microbes and releasing cytokines and other chemical messengers that trigger a full immune response. Helper T cells, or CD4+ T cells, use cytokine signaling to activate surrounding cells, exponentially increasing the number of immune cells attacking a pathogen. Helper T cells can also activate B cells to create and release specific antibodies designed to destroy microbes. 

CD8+ T cells, or cytotoxic killer T cells, are also activated during an inflammatory cascade and directly attack and eliminate pathogenic or infected cells. Regulatory T cells limit the actions of the other immune cells, thus preventing harm from over-inflammation. 

While effector T cells are often more robust immune cells than naive T cells, all T cells present their specific type of fragility during cell separation and processing. T cells are sensitive to exposure to harsh environments that do not meet their environmental requirements—such as optimal pH—and can undergo activation-induced cell death (AICD) when excessively stimulated. Appropriate handling must be followed during cell isolation, culturing, and separation to maintain stimulation levels and avoid AICD. 

B Cells

B lymphocytes are another important cell in the immune system, responsible for antibody production and adaptive immunity. When B cells are excessively disturbed due to rough handling, experience prolonged changes in their environment, or are subjected to excess stress, they are prone to damage or cell death. They must be maintained with the proper culture media, culture time, growth factors, and cytokines for survival. 

Improper Handling Affects Downstream Results

Prolonged exposure to stressful conditions or suboptimal environments can have detrimental effects on the population of cells within a culture or their performance post-separation in downstream assays.

Stressful conditions and harsh environments can induce genetic changes within a cell that lead to upregulation or downregulation of the genes essential to the function of each cell type. This change in regulation and expression can result in unreliable downstream assays. 

Improper handling leading to stress can lead to the breakdown or inhibition of cell communication pathways. Blocked pathways potentially affect cell proliferation, differentiation, apoptosis, and inflammatory responses. 

Rough handling and stress change cell morphology. In some immune cells, morphology plays a major role in determining cell function, as surface enzymes and structures allow the cell to interact with its surroundings. A change in cell morphology could cause a cell to behave atypically in downstream tests, such as experiencing difficulties taking in nutrients, removing waste from the cellular matrix, or communicating with other cells. 

Apoptosis is a typical cell response to stress and leads to decreased cell viability in cultures overall. The loss of a significant portion of a cell culture to apoptosis can skew the population distribution and negatively affect assay results. 

Stress is a common trigger of an inflammatory response. Once started, the activation transfers between neighboring cells exponentially, causing widespread inflammation. Inflammatory signals cause changes in gene expression and cellular responses, jeopardizing experimental outcomes including protein expression and cell functionality. 

How to Optimize Cell Handling

Avoiding roughness in rare cell isolation can make a significant difference in the viability and experimental success of cell cultures. Gentle handling includes slow and controlled pipetting, as well as avoiding unnecessary vortexing, shaking, or centrifugation. 

Cells should not be outside of their preferred environment for long periods. Timely and organized experimentation ensures proper handling and maintenance. Keep in mind all cell types have different and unique culture requirements to survive, grow, and function. By finding the appropriate media, pH, temperature, and humidity, the cell population can thrive for much longer. 

Microbubbles for Gentle Cell Sorting

Comparing the available cell separation methods to determine the best solution for each type of cell also helps protect viable cells. Akadeum provides an unmatched gentle cell-sorting solution available, ensuring minimal cell loss while isolating unique cell populations from damage during cell separation and protecting the samples from degradation and apoptosis. 

By utilizing natural buoyancy for negative selection, our microbubbles gently sift through cells, targeting unwanted contaminants or cell types with antibodies, meaning they simply float to the top of the sample where they can be removed. Avoid harsh lengthy clean-up procedures that cost valuable cells and time by skipping the magnets and columns and using microbubbles instead. 

Akadeum’s microbubble technology can also be utilized to clean up previously separated or damaged cell samples to regain cell population purity using the Dead Cell Removal Kit. Our negative selection buoyancy-activated cell sorting microbubbles provide a gentle method for isolating truly untouched cells of interest for the most accurate downstream results.