Levitating Cells to Sort the Fit and the Fat

Density is a core material property and varies between different cell types, mainly based on differences in their lipid content. Sorting based on density enables various biomedical applications such as multi‐omics in precision medicine and regenerative repair in medicine. However, a significant challenge is sorting cells of the same type based on density differences. Here, a new method for real‐time monitoring and sorting of single cells based on their inherent levitation profiles driven by their lipid content is reported. As a model system, human‐induced pluripotent stem cell (hiPSC)‐derived cardiomyocytes (CMs) from a patient with neutral lipid storage disease (NLSD) due to loss of function of adipose triglyceride lipase (ATGL) resulting in abnormal lipid storage in cardiac muscle are used. This levitation‐based strategy detects subpopulations within ATGL‐deficient hiPSC‐CMs with heterogenous lipid content, equilibrating at different levitation heights due to small density differences. In addition, sorting of these differentially levitating subpopulations are monitored in real time. Using this approach, sorted healthy and diseased hiPSC‐CMs maintain viability and function. Pixel‐tracking technologies show differences in contraction between NLSD and healthy hiPSC‐CMs. Overall, this is a unique approach to separate diseased cell populations based on their intracellular lipid content that cannot be achieved using traditional flow cytometry techniques.

[1]  H. Mizumoto,et al.  Expansion and differentiation of human iPS cells in a three-dimensional culture using hollow fibers and separation of the specific population by magnetic-activated cell sorting. , 2019, Journal of bioscience and bioengineering.

[2]  Matthew Miyamoto,et al.  Large particle fluorescence-activated cell sorting enables high quality single cell RNA-sequencing and functional analysis of adult cardiomyocytes , 2019, bioRxiv.

[3]  S. M. Chambers,et al.  Lipid Deprivation Induces a Stable, Naive-to-Primed Intermediate State of Pluripotency in Human PSCs. , 2019, Cell stem cell.

[4]  K. Shimada,et al.  Triglyceride deposit cardiomyovasculopathy: a rare cardiovascular disorder , 2019, Orphanet Journal of Rare Diseases.

[5]  M. Schweiger,et al.  Of mice and men: The physiological role of adipose triglyceride lipase (ATGL)☆ , 2019, Biochimica et biophysica acta. Molecular and cell biology of lipids.

[6]  W. Tan,et al.  Following hearts, one cell at a time: recent applications of single-cell RNA sequencing to the understanding of heart disease , 2018, Nature Communications.

[7]  H. Tojo,et al.  Generation of Fabry cardiomyopathy model for drug screening using induced pluripotent stem cell-derived cardiomyocytes from a female Fabry patient. , 2018, Journal of molecular and cellular cardiology.

[8]  Ahu Arslan Yildiz,et al.  Biofabrication of in situ Self Assembled 3D Cell Cultures in a Weightlessness Environment Generated using Magnetic Levitation , 2018, Scientific Reports.

[9]  F. Xiao,et al.  Patients with neutral lipid storage disease with myopathy (NLSDM) in Southwestern China , 2018, Clinical Neurology and Neurosurgery.

[10]  S. Kelley,et al.  Amplified Micromagnetic Field Gradients Enable High-Resolution Profiling of Rare Cell Subpopulations. , 2017, ACS applied materials & interfaces.

[11]  Y. Yoon,et al.  Current Strategies and Challenges for Purification of Cardiomyocytes Derived from Human Pluripotent Stem Cells , 2017, Theranostics.

[12]  R. Coleman,et al.  Generation of induced Pluripotent Stem Cells as disease modelling of NLSDM , 2017, Molecular genetics and metabolism.

[13]  K. Drosatos,et al.  Lipid Use and Misuse by the Heart. , 2016, Circulation research.

[14]  Naside Gozde Durmus,et al.  Magnetic levitation of single cells , 2015, Proceedings of the National Academy of Sciences.

[15]  M. Radisic,et al.  Enrichment of live unlabelled cardiomyocytes from heterogeneous cell populations using manipulation of cell settling velocity by magnetic field. , 2013, Biomicrofluidics.

[16]  Luke P. Lee,et al.  Label-free electrophysiological cytometry for stem cell-derived cardiomyocyte clusters. , 2013, Lab on a chip.

[17]  Sean P. Palecek,et al.  Directed cardiomyocyte differentiation from human pluripotent stem cells by modulating Wnt/β-catenin signaling under fully defined conditions , 2012, Nature Protocols.

[18]  Boyang Zhang,et al.  Label-Free Enrichment of Functional Cardiomyocytes Using Microfluidic Deterministic Lateral Flow Displacement , 2012, PloS one.

[19]  R. Zechner,et al.  Neutral lipid storage disease: genetic disorders caused by mutations in adipose triglyceride lipase/PNPLA2 or CGI-58/ABHD5. , 2009, American journal of physiology. Endocrinology and metabolism.

[20]  J. Menéndez,et al.  Fatty acid synthase and the lipogenic phenotype in cancer pathogenesis , 2007, Nature Reviews Cancer.

[21]  M. Lathrop,et al.  The gene encoding adipose triglyceride lipase (PNPLA2) is mutated in neutral lipid storage disease with myopathy , 2007, Nature Genetics.

[22]  Nicole Pamme,et al.  Magnetism and microfluidics. , 2006, Lab on a chip.

[23]  Y. Lo,et al.  Increased serum lipids are associated with higher CD4 lymphocyte count in HIV‐infected women , 2006, HIV medicine.

[24]  Mehmet Toner,et al.  Size-based microfluidic enrichment of neonatal rat cardiac cell populations , 2006, Biomedical microdevices.

[25]  E. Wagner,et al.  Defective Lipolysis and Altered Energy Metabolism in Mice Lacking Adipose Triglyceride Lipase , 2006, Science.

[26]  J P Clarys,et al.  Adipose tissue density, estimated adipose lipid fraction and whole body adiposity in male cadavers. , 1994, International journal of obesity and related metabolic disorders : journal of the International Association for the Study of Obesity.

[27]  L. Sjöström,et al.  Isolation and characterization of cells from rat adipose tissue developing into adipocytes. , 1978, Journal of lipid research.