The alarming rate at which obesity is growing worldwide has intensified the necessity and urgency of obtaining a deeper understanding of all aspects of adipocyte biology, as obesity often leads to numerous chronic and costly health conditions. Substantial discoveries have been made in the complex functions of adipocytes that have led to
advances in endocrinology, cardiology, energy metabolism, cancer biology, and developmental and stem cell biology. Scientists continue to study the implications of managing adipocyte growth and proliferation in a variety of disease processes to uncover opportunities for unique therapeutic interventions.
The ongoing quest for insight into the varied roles and mechanisms of these cell types puts adipocyte research at the heart of a variety of diseases such as diabetes, hypertension, heart disease, metabolic syndrome, sleep disorders, and cancer. As such, the need to isolate and characterize adipocytes is immense.
However, adipocytes are notoriously problematic to work with. These highly sensitive large cells tend to form bulky clumps that make them challenging to isolate using conventional separation techniques such as FACS. The degree of differentiation in precursor models also poses considerable challenges in assessment and downstream applications due to the limitations in current techniques that leave an abundance of undifferentiated cells in the samples. The LeviCellTM provides a solution to these limitations.
The LeviCellTM platform separates cells based on physical properties such as density. Adipocytes have lower density compared to most mammalian cells due to their lipid-rich vesicles causing them to levitate higher in the LeviCellTM environment. The nature of the LeviCellTM technology enables robust enrichment, label-free isolation, and minimal cell loss and damage of these delicate adipocytes.
Since the entire process occurs in a completely closed environment, prep time, contamination potential, and cellular damage from repeated handling are minimized. Furthermore, the gentle processing method of the LeviCellTM, which uses <1 psi of pressure, accommodates the safe and effective isolation of large cell clumps (up to 400µm) – ensuring maximum viability, even with these highly sensitive cells.
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Sample pipetted into the LeviCell™ cartridge.
Automated Label-Free Sorting
Precision density gradients drive levitation based on cells physical properties.
Selected populations collected into separate ports for further analysis or use.
Undifferentiated precursor cells levitate at a uniform height, which indicates a homogeneous cell population. These cells were suspended in 100 mM Levitation Agent and allowed to reach their final levitation height shown in Figure 1a.
Precursor cells differentiated for 10 days reveal 2 distinct populations. The top layer of cells indicate differentiated adipocytes, whereas the lower layer contains the undifferentiated cells. The dark clumps at the lower portion of the top layer are collections of cells, demonstrating the ability of the LeviCellTM to process large clusters of cells
Cell Preparation for LeviCellTM
Different concentrations of Levitation Agent were used to determine which would provide maximum levitation distance between the two adipocyte cell populations.
Maximum levitation height difference between differentiated and undifferentiated adipocytes was achieved using 30 mM of Levitation Agent (Figure a above). Note that as the concentration of Levitation Agent increased, overall levitation height of both populations increased while the distance between differentiated and undifferentiated cells decreased
Starting samples for Fig c,d were differentiated for 7 days, indicating that the LeviCellTM can effectively isolate and enrich sensitive cells such as adipocytes after relatively early differentiation time points.
The LeviCellTM‘s demonstrated ability to efficiently and effectively isolate large, fragile adipocytes in a gentle, label-free, closed environment shows the platform’s immense potential to reset the bar in cell separation technology. Significant advances in understanding adipogenesis, adipocyte-related diseases, and fat tissue engineering are within reach as the LeviCellTM unambiguously exhibits its ability to enrich an abundance of pure and viable adipocytes in a shorter time span than conventional techniques.