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    CNS top issue!

    • Last Update: 2022-02-22
    • Source: Internet
    • Author: User
    Search more information of high quality chemicals, good prices and reliable suppliers, visit www.echemi.com
    The human brain is actually extremely complex.
    The brain that only occupies a small part of our body contains hundreds of billions of neurons, and there are elusive and complex connections between them, but this does not prevent scientists from being stunned by her mystery.
    attracted
    .

    Faced with such a "precise" organ, how to efficiently conduct multi-faceted research on it has always been a headache for many scientists: 1) Popular methods such as immunostaining or flow cytometry (FACS) 2) key stem cell and neural progenitor cell populations are difficult to isolate without prior knowledge of specific molecular markers; 3) early single-cell RNA-sequencing techniques (scRNA-seq) is extremely limited by low cell numbers
    .

    The birth of the BD Rhapsody single-cell multi-omics analysis system can easily solve the above technical problems in brain research
    .

    1) The system is currently the only single-cell platform that can obtain transcriptome, proteome, immune panel (VDJ) and multi-sample analysis results in the same experiment.
    The rich multi-dimensional parameters make the research of brain science more in-depth and comprehensive.
    ;2) BD Rhapsody's unique DNA molecular tags include as many as 880,000 cell labels and about 7.
    5 million gene tags (Unique Molecular Index).
    Precise labeling and efficient isolation of stem cells and neural progenitor cell populations; 3) Compared with earlier low-throughput single-cell RNA sequencing methods, BD Rhapsody has higher cell throughput and sensitivity.
    based) principle, 100-40,000 cells can be isolated in a single experiment, and the platform’s requirement for cell suspension concentration can be as low as 40 cells/ul, which is also applicable to some trace samples in brain research, such as cerebrospinal fluid
    .

    Because of this, "BD Rhapsody Single-Cell Multi-omics Analysis System" has become the best partner for "brain/neural" single-cell research, helping scientists publish many high-scoring papers
    .

    This article selects several of the most representative literatures in recent years and reviews them with you - the development of No.
    1 human cerebral cortex involves the migration and integration of neurons
    .

    A research team from Stanford University used a 3D differentiation method to successfully induce the directional development of neurospheres (neural spheroids) into cerebral cortical spheroids (human cortical spheroids, hCS) and subcortical spheroids (human subpallium spheroids) in a petri dish , hSS) and assembled the two in vitro to facilitate cell migration and development of human cortical circuits
    .

    To comprehensively characterize the transcriptomic information of hCS and hSS single cells, the researchers used BD Rhapsody to combine and enzymatically dissociate 11,838 hCS or hSS into single cells on day 105 of their differentiation for batch transcriptome sequencing analysis
    .

    Among them, single cell capture is achieved by random distribution of single cell suspensions in >200,000 microwells by limiting dilution
    .

    Subsequently, the researchers prepared whole transcriptome libraries according to the BD Rhapsody single-cellwhole-transcriptome amplification workflow, and used the BD Rhapsody analysis pipeline to process the analysis data
    .

    Clusters of cells isolated from hCS or hSS by the t-distributed stochastic neighborhood embedding (tSNE) method showed the separation under both conditions
    .

    Electrophysiological tests of the assembled cells showed that GABAergic and glutamatergic neurons were able to successfully form circuits and send signals to each other
    .

    This achievement undoubtedly provides a useful model for studying neurodevelopment and disease, as well as neural circuits
    .

    No.
    2 In order to verify the reliability of in vitro differentiation of specific organs, researchers from Stanford University in the United States cultured and induced 12 human induced pluripotent stem cell lines (hiPSC lines) to differentiate into hCSs in vitro.
    BD Rhapsody performed single-cell transcriptome analysis of differentiation products to confirm the reproducibility of the process: On day 105 of differentiation, we differentiated three parallels into hCS-FF (Feeder-Free condition, trophoblast-free condition) ) of hiPSC lines were digested into single cells and captured
    .

    Then, by sub-sampling BD Rhapsody magnetic beads, a whole transcriptome library is prepared from about 67% of the captured cells, followed by cDNA synthesis and other operations, and the BD Rhapsody analysis pipeline is used to process the sequencing data to determine cell markers and molecules index
    .

    Finally, single-cell transcriptome profiles were analyzed using BD Data View and the current data results were compared with hCS-MEF (Mouse Embryonic Fibroblasts, mouse embryonic fibroblasts) and hSS single-cell data previously acquired on the same platform (n=24,237)
    .

    tSNE cluster analysis found that clustering of cell populations from hCS and hSS revealed differences in dorsal and ventral differentiation states
    .

    This study shows that the experimental protocol for culturing neural stem cells in vitro has good repeatability between different experimental conditions and different stem cell lines, and the differences in cell transcriptomes are mainly derived from the differentiation stage, not the cell lineage or experimental conditions
    .

    No.
    3 Fibroblast growth factor 21 (FGF21) is a liver-derived hormone that signals glutamatergic neurons in the ventromedial hypothalamus (VMH) to inhibit carbohydrates intake
    .

    In this trial, a research team from the University of Iowa School of Medicine and the University of Copenhagen, Denmark, took advantage of the BD Rhapsody single-cell sequencing platform to rationally assess which cells express FGF21's coreceptor KIb (β-klotho) in the hypothalamus.
    ): First, target single cells in mouse hypothalamus were screened by BD FACS AriaII, and then the screened cells were loaded onto BD Rhapsody to capture single cells, and cDNA was synthesized
    .

    Subsequently, a cDNA library was generated using two custom primer panels, which were designed by BD and contained low-abundance expressed genes and high-abundance expressed genes, respectively
    .

    Using this method, along with the BD Resolve analysis pipeline, the researchers performed efficient cell sorting, quality filtering, and sequencing data annotation on live cells, resulting in 1,904 sorted putative Klb cells
    .

    Subsequently, using tSNE analysis and unsupervised clustering, clusters of 12 neuronal and non-neuronal cell types were identified
    .

    Through this pathway, the researchers were able to precisely identify FGF21 target cells in the hypothalamus and revealed that activation of FGF21 signaling in glutamatergic neurons is necessary and sufficient to mediate FGF21-induced sugar suppression and sweet taste preference
    .

    No.
    4 The research group of Professor Liu Qiang from Tianjin Medical University and Tianjin Institute of Neurology reported in the previous literature the immune mechanism of intracerebral hemorrhage (ICH) formation after cerebral hemorrhage (Perihematomal edema, PHE)
    .

    During the experiment, it uses BD Rhapsody to capture single cells, and uses BD Rhapsody WTA Amplification Kit to construct single-cell RNA-seq library, uses BD Rhapsody data analysis process to process sequencing data, and uses BD Data View to analyze single-cell transcriptome profiles , plotted the gene expression of 12,443, 8,751 and 4,270 natural killer (NK) cells in the blood control (sham blood), ICH blood and ICH brain samples spectrum
    .

    Among them, NK cells were isolated from spleen cells of naive C57BL/6 mice (BD Bioscience), purified and subjected to two rounds of cell sorting on the FACS Aria III system
    .

    Single-cell sequencing and immunological studies revealed that NK cells were the predominant immune cell subset in the early stages of ICH, and unbiased clustering of single-cell transcriptional profiles revealed that after intracerebral hemorrhage, the two major NK cell subsets had high levels of cells, respectively.
    Toxic or potent chemokine production characteristics, distinguishable from surrounding NK cells
    .

    BD Rhapsody single-cell sequencing technology is playing an irreplaceable role in the field of brain science
    .

    With this tool, many morphogenetically and functionally distinct brain cell types have been identified, giving us a better understanding of the diversity of cell types in the brain, the specific functions or states of cells, etc.
    know
    .

    Main highlights of BD Rhapsody™ single-cell multi-omics analysis system: High sample compatibility: Powerful Microwell-based technology and BD Medical's mature flow sorting methods make single-cell experiments no longer affected by sample types.
    Multi-omics: a rare single-cell analysis platform that can do transcriptome, immunology and proteome at the same time; high efficiency and low cost: high-efficiency cell capture rate, support 1-12 samples synchronously loading, synchronously constructing library to effectively reduce batch-to-batch differences; high quality: Scanner visual quality control system to ensure experimental quality and reduce experimental losses; high-quality service platform: provide a series of customized chemical bioinformatics analysis programs, supporting bioinformatics software and professional bioinformatics support Team, Shengxin Xiaobai can also easily get started
    .

    Want to get the original text of the above literature? Follow the BD Rhapsody single-cell platform and fill in the questionnaire to get it.
    How to fill in the questionnaire: Ø Scan the code to follow the official account, and click to complete the questionnaire; Ø For old users, please send the [questionnaire] on the official account, and click the link to fill in the questionnaire.
    Contact Manager Liu: 15312997988, learn more about BD Rhapsody single-cell multi-omics analysis system Reference: 1.
    Birey, F.
    et al.
    Assembly of functionally integrated human forebrain spheroids.
    Nature 545,54–59 (2017).
    https://doi.
    org/10.
    1038/nature223302.
    Yoon, S.
    -J.
    et al.
    Reliability of human cortical organoid generation.
    Nature Methods 16, 75–78 (2019).
    https://doi.
    org/10.
    1038/s41592-018-0255- 03.
    Jensen-Cody, SO etal.
    FGF21 Signals to Glutamatergic Neurons in the Ventromedial Hypothalamusto Suppress Carbohydrate Intake.
    Cell Metabolism 32, 273-286.
    e6 (2020).
    https://doi.
    org/10.
    1016/j.
    cmet.
    2020.
    06 .
    0084.
    Li, Z.
    et al.
    Braintransforms natural killer cells that exacerbate brain edema after intracerebralhemorrhage.
    Journal of Experimental Medicine 217, e20200213 (2020).
    https://doi.
    org/10.
    1084/jem.
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