The natural killer cells are a type of lymphocytes, primarily involved in regulation of the immune system. They are called “Natural” because they don’t require prior activation in order to recognize abnormal and/or infected cells to lyse them. These cells are also able to recognize and kill cells that have downregulated or lost major histocompatibility […]
Available Format: ≥1000000 cells cryopreserved in suitable medium along with DMSO.
The natural killer cells are a type of lymphocytes, primarily involved in regulation of the immune system. They are called “Natural” because they don’t require prior activation in order to recognize abnormal and/or infected cells to lyse them. These cells are also able to recognize and kill cells that have downregulated or lost major histocompatibility complex (MHC) on their surface, which can occur in cancer cells and/or virus infected cells. These cells are also involved in secretion of cytokines, which are signalling molecules that help to coordinate and regulate immune response. These cells are found in the blood, lymph nodes and spleen.In humans, NK cells represent around 8-20% of the circulating lymphocytes. Unlike other lymphocytes, NK cells do not express antigen-specific receptors. NK cells play a critically important role in regulating adaptive immune response. Studies are ongoing to understand the role of NK cells in activation of cytokines post viral infections. Due to their ability to recognize tumor cells, NK cells are widely studied as primary components in immune-oncology.Kosheeka has validated isolation protocol for peripheral blood derived NK cells using positive immunomagnetic separation. However, we have also validated other methods of isolation of NK cells like differential centrifugation or fluorescence activated cell sorting; depending upon experimental demands. Kosheeka NK cells are traditionally characterized for the presence of CD56 cell surface markers and the absence of CD3. Due to separation of cells through immunomagnetic selection, the isolated cells express highest purification. All the donors are tested for infectious markers.
Product Category
Homo Sapiens, Human
Product Type
Adherent cultures, suspension cell population
Derived From
Peripheral Blood
Cell Morphology
Small-to-medium sized lymphocytes with large granular cytoplasm and a round nucleus.
Culture & Growth Properties
Adherent, proliferating population
Passage No.
Customized
Mycoplasma
Detected on demand
Hepatitis B
Negative
Hepatitis C
Negative
HIV-1
Negative
Aerobic & Anaerobic Bacteria
Negative after 7 days of culture
Positive for
CD 56, CD16
Negative for
CD 3
NK cells are isolated from peripheral blood of a healthy donor with the help of apheresis. The commonly used techniques for the isolation of NK cells from peripheral blood are differential centrifugation using lymphoprep, positive/negative selection through application of magnetic field, or fluorescence activated cell sorting. Once NK cells are isolated, they can further be purified, activated, expanded in vitro for use in research and development or in therapeutics as well.
It should also be noted that the purity and yield of isolated NK cells depends largely on the employment of suitable methods of isolation. Kosheeka recommends the validation of isolation method of NK cells before using them in experiments or in therapeutics.
Yes, it is possible to expand NK cells in culture without inducing differentiation. In fact, many studies have confirmed expansion and activation of NK cells using a set of cytokines and/or cocktail of growth factors, promoting their activation and proliferation without subjecting them to differentiation.
One of the common approaches to expand NK cells in culture is to use cytokines such as IL-2, IL-15, and IL-21. These interleukins are widely accepted in different researches to promote NK cell survival and proliferation. These cytokines can also be added to the culture medium, along with other factors such as using feeder cells to support expansion of NK cells.
Recently, certain studies have also promoted use of artificial antigen-presenting cells (APCs) that express specific ligands for activating receptors on NK cells. These APCs are responsible for stimulating the expansion of NK cells without inducing any differentiation.
It is important to know that the expansion and activation of NK cells proliferation can affect their phenotypic and genotypic functioning. Thus, the optimal conditions required to expand NK cells can vary depending upon experimental requirements. Therefore, it is important to validate the requirements before using them for research or other applications.
Although, the choice of culture media is largely dependent upon the applications and experimental requirements; in general RPMI160, lymphocyte separation media and serum free medium is used along with autologous human serum to expand NK cells in culture without inducing any differentiation. However, Kosheeka emphasizes on validation of protocol before routine implementation of protocol for further optimization.
Several methods can be employed to quantify NK cells, the most common amongst them are flow cytometry, ELISA, cell counting etc.
NK cells are characterized based on their morphology, phenotypic specifications and functional contribution. More commonly, NK cells are characterized based on their morphology like size, shape and staining properties. Based on the presence of specific surface markers, NK cells are characterized using flow cytometry and immunohistochemistry; typically for the expression of certain markers like CD56, CD16, in addition to CD3, and CD8 depending upon the activation state and functional specialization. Based on the functional properties, NK cells can further be characterized for their cytotoxicity, cytokine secretion, and immunoregulatory activity. In addition to the same, gene expression profiling and epigenetic analysis can provide insights into the molecular mechanisms that underlie the functional properties of NK cells.
The lifespan of NK cells in the culture can vary depending upon number of factors like culture conditions, the source of NK cells, along with the activation state of the cells. Generally, NK cells can be sustained in the culture for several weeks to a few months, although their viability and function may decline over the period of time.
The culture conditions like the medium, cytokines and other supplements can affect the survival and proliferation of NK cells . Additionally, certain studies have specified that culture conditions like the use of feeder cells can provide signals that support the growth and survival of NK cells. The source of NK cells can also affect their lifespan in culture, like the NK cells isolated from bone marrow and/or peripheral blood can affect their survival and proliferation, due to replicative senescence or activation induced cell death.
Overall, the lifespan of NK cells in culture can be expanded by optimizing the culture conditions, using appropriate sources of NK cells, and carefully controlling their activation state.
Depending upon the choice and demands, NK cells can be transported through several standard methods like cryopreserved cells at ultra-low temperature, refrigerated temperature for a short period of time (up to 72 hrs) using specialized transport containers, further maintaining stable temperature. The NK cells can also be transported in cell culture medium with appropriate temperature and a cocktail of nutrients and other supplementation for optimum growth.
Yes, the expansion of NK cells in culture typically requires the addition of a cocktail of cytokines and other growth factors to promote cell survival and proliferation. The most commonly used cytokines for NK cell expansion are interleukin 2, IL-15 and IL-21. In addition to cytokines, the use of feeder cells, such as irradiated PBMCs or K562 cells can provide additional signals that support the growth and survival of NK cells in culture. The feeder cells can also provide important growth factors and cytokines that promote NK cells proliferation and survival.
Yes, human serum can be used for the expansion of NK cells in the culture, supporting their optimal growth and proliferation. It contains a complex mixture of cytokines and other growth factors, responsible for supporting proliferation of NK cells. However, it should also be noted that batch variability of the human serum may directly impact growth and proliferation rate of NK cells. Thus, it is always recommended to get human serum of a reputed brand.
Yes, we do supply human serum for the expansion of NK cells. Please drop a mail to us at info@kosheeka.com or you can directly call us/WhatsApp us at +919654321400 and our scientific team will get back to you at the earliest.
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