Application of single cell suspension dissociators in 4 types of experiments on cells
| December 1, 2023
Cells are the basic building blocks of all living organisms. This is why most biological research focuses on cells. Cells can be cultured either by primary cells isolated directly from tissues or by cell lines. Isolating single cells from tissues for primary cell culture is commonly seen. These primary cells will be subsequently used by experimenters in various scenarios. However, there remain a number of limitations in obtaining single cells from parenchymal tissues in the laboratory, including incomplete digestion, mediocre viability, and destruction of cell-surface antigens.
Single cell suspension dissociators reduces manual manipulation and greatly improved the efficiency of single cell suspension preparation. Their gentle procedures are conducive to keeping surface antigens and enhancing cell viability. Specific application scenarios are as follows:
1. Organoid culture
Organoid culture is a tissue culture method to grow functional 3D organoids from a group of cells and combination of various biochemical factors. It has the tissue structure and function of a real organ. It can be applied in organ research, drug screening, and treatment by simulating cellular interactions and physiological processes within the organ. Typically, cells are extracted from tissues, and then cultured in specialized media. Single cell suspension dissociators play a vital role during cell extraction, improving cell yield and viability, thereby increasing organoid survival probability. Organoids can self-organize in three dimensions, which express specific cell markers and functional proteins. In addition, by adjusting culture conditions, such as oxygen level, room temperature, and cell density, the physiological environment of the organ can be better simulated.
2. Cell sorting
Cell sorting is the process through which a particular cell type is separated from others and then cultivated in vitro. It can be used to purify specific cell populations, study their properties and functions, carry out related experiments, and explore applications. Cell suspension preparation, a prerequisite for cell sorting, can be realized via single cell suspension dissociators. At present, methods of cell sorting fall into two major categories: fluorescence activated cell sorting (FACS) and magnetic-activated cell sorting (MACS). Based on particular surface markers or biological properties of cells, these techniques use specific antibodies or molecular probes to distinguish target cells from others. After sorting, target cells are collected individually and cultured in media or bioreactors. Cell sorting brings about purified cell populations, eliminating the interference of other cells on experimental results, so that characteristics and functions of target cells can be better studied and understood. Furthermore, cell sorting enables enough target cells for future experiments and applications.
3. Single-cell sequencing
Single-cell sequencing is a high-resolution genomics technology used for comprehensive analysis of gene expression in individual cells. Compared with traditional sequencing that examines the genome of a homogeneous cell population, single-cell sequencing can reveal cellular heterogeneity in cell populations and functional differences in individual cells. Through single-cell sequencing, researchers can identify and classify different types of cells, studying how they differentiate, develop, and respond in various environments or disease states. In addition, this technology also reveals the interactions and communication networks among cells, helping us better understand the functions and regulatory mechanisms of cellular systems. Processes of single-cell sequencing usually include isolation of single cells, cytolysis and RNA extraction, cDNA synthesis and library construction, and high-throughput sequencing. The premise for these steps is to successfully dissociate the tissue sample into individual cells. Single-cell suspension dissociators efficiently convert tissues into single-cell suspensions, providing high-quality samples for subsequent sequencing.
4. Tissue homogenization
Tissue homogenates, a uniform mixture converted by disrupted biological tissues, are used for determining cytoplasmic and nuclear contents, extracting enzymes and other cellular components, and conducting other biological research. Processes of tissue homogenization usually involve mincing/grinding tissue samples, adding enzymes/proteases to appropriate buffers, and digesting tissues and releasing cells. All these can be completed with a single-cell suspension dissociator. Subsequently, tissue homogenates are obtained after centrifuging or filtering samples to remove undigested fragments. These homogenates can be applied in isolating intracellular components such as organelles, nucleic acids, and proteins.
In general, single cell suspension dissociators are widely used in various cell experiments. Featuring high-efficiency and automation, they make cell sample preparation more convenient and reliable, providing essential support for cytological research, biomedical study and clinical applications. With technological advancement, single cell suspension dissociators will play an increasingly vital role in research and applications in cytology.