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Human plasma-like medium improves T lymphocyte activation

ABSTRACT: Human plasma-like medium improves T lymphocyte activation

PROVIDER: PRJNA560687 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR9993430_1.fastq.gz	Fastqsanger.gz
	SRR9993430_2.fastq.gz	Fastqsanger.gz
	SRR9993431_1.fastq.gz	Fastqsanger.gz
	SRR9993431_2.fastq.gz	Fastqsanger.gz
	SRR9993432_1.fastq.gz	Fastqsanger.gz

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Human plasma-like medium improves T lymphocyte activation

Project description:T lymphocytes are critical effectors of the immune system and the ability to study their behavior in synthetic media in vitro has facilitated major discoveries in their development, function, and fate. Recently, it has been shown that the constituents in human plasma differ markedly from commonly used synthetic media and these differences have important effects on cell physiology. We therefore compared T lymphocyte activation in human plasma-like medium (HPLM) to RPMI conventional medium both with and without serum. We found that HPLM and RPMI induced vastly different transcriptional programs, especially for transcripts encoding proteins involved in activation, proliferation and metabolism, in human peripheral blood T cells after T cell receptor (TCR) stimulation. We also found that in dialyzed fetal bovine serum (dFBS), HPLM supports a stronger response to TCR engagement compared to RPMI. In reconstruction experiments, we demonstrated that this activation difference is due to RPMI being hypocalcemic relative to the in vivo milieu and that addition of calcium chloride to RPMI can improve human T lymphocyte activation. Thus, we conclude that investigators should be cognizant of differences between commonly used media formulations and the in vivo environment since this could profoundly affect their experimental results and that physiologic media is a valuable new way to study immune cells in culture

2020-01-21 | GSE135936 | GEO

Human Plasma-like Medium Improves T Lymphocyte Activation.

Project description:T lymphocytes are critical for effective immunity, and the ability to study their behavior in vitro can facilitate major insights into their development, function, and fate. However, the composition of human plasma differs from conventional media, and we hypothesized that such differences could impact immune cell physiology. Here, we showed that relative to the medium typically used to culture lymphocytes (RPMI), a physiologic medium (human plasma-like medium; HPLM) induced markedly different transcriptional responses in human primary T cells and in addition, improved their activation upon antigen stimulation. We found that this medium-dependent effect on T cell activation is linked to Ca2+, which is six-fold higher in HPLM than in RPMI. Thus, a medium that more closely resembles human plasma has striking effects on T cell biology, further demonstrates that medium composition can profoundly affect experimental results, and broadly suggests that physiologic media may offer a valuable way to study cultured immune cells.

| S-EPMC6941860 | biostudies-literature

Comparison of the lymphocyte before and after the activation.

2019-08-31 | GSE86293 | GEO

Pharmacokinetics and Metabolic Activation of Capecitabine

Project description:The objective of this pharmacokinetic study is to exclude a possible influence of CETUX on the plasma disposition and metabolic activation of Capecitabine (CCB) and when this regimen is given combined with Oxaliplatin (OxPt).

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Transcriptome and chromatin accessibility in oral squamous carcinoma with human plasma-like medium culture (RNA-Seq)

Project description:In recent years, human plasma like medium (HPLM) have been developed that mimic the conditions of the human physiology. In comparison with conventional cell culture medium, cell culture in HPLM had extensive effects on cellular metabolism, including metabolome, redox status, and glucose utilization. However, molecular mechanisms underlying HPLM effects on human cells are only partly understood. Transcriptome and chromatin accessibilities are important regulators of cell states and implicate in a variety of biological processes. We habituated three oral cancer cell lines such as SAS, HSC-3, and HSC-4 in HPLM and evaluated cell growth and drug sensitivity. We perform transcriptome analysis between HPLM and conventional culture condition in HSC-3 cells by mRNA-seq.

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Transcriptome and chromatin accessibility in oral squamous carcinoma with human plasma-like medium culture (ATAC-Seq)

Project description:In recent years, human plasma like medium (HPLM) have been developed that mimic the conditions of the human physiology. In comparison with conventional cell culture medium, cell culture in HPLM had extensive effects on cellular metabolism, including metabolome, redox status, and glucose utilization. However, molecular mechanisms underlying HPLM effects on human cells are only partly understood. Transcriptome and chromatin accessibilities are important regulators of cell states and implicate in a variety of biological processes. We habituated three oral cancer cell lines such as SAS, HSC-3, and HSC-4 in HPLM and evaluated cell growth and drug sensitivity. We perform epigenetic analysis between HPLM and conventional culture condition in HSC-3 cells by ATAC-seq.

2024-02-21 | GSE256192 | GEO

B lymphocyte activation induces DNA methylation reprogramming and establishes a common epigenetic signature in memory B and plasma cell compartments [DNA methylation profiling]

Project description:DNA methylation profiling of human B cell populations representing a developmental series before and after immune activation. The B cells are derived from inflamed tonsils. The B cells were already activated in vivo in patients, therefore there was no need to stimulate the B cells after isolation. Gene expression profiling was also performed from the same samples. Four B cell subsets: Naïve, germinal center B cells (GC), memory B cells, and plasma cells (PC) were purified by FACS ex vivo from tonsils of 8 human subjects. Methylated CpG island Recovery assay (MIRA) was utilized to enrich for methylated DNA fragments from each sample for microarray analysis. *** This Series reports DNA methylation profiling data. ***

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Project description:Immunotherapy using activated lymphocytes has recently been developed, but there is not yet the biomarker which can predict curative effect. In our recent study, we found that random migration is significantly increased in activated lymphocyte. Furthermore, it is suggested that random migration may affect cytotoxicity. Therefore we considered that migration of lymphocyte may become the effect prediction biomarker of Immunotherapy. And then we analyzed the gene which fluctuated by lymphocyte activation to search the molecules which regulate migration of lymphocyte.As the result, we found out that expression of PTPN3 fluctuates before and after activation of the lymphocyte.Next, we compared the gene expression of the activated lymphocyte into which shPTPN3 was introduced and of the activated lymphocyte into which a control vector was introduced to search the action mechanism of PTPN3.

2019-10-31 | GSE89328 | GEO

B lymphocyte activation induces DNA methylation reprogramming and establishes a common epigenetic signature in memory B and plasma cell compartments [gene expression profiling]

Project description:DNA methylation profiling of human B cell populations representing a developmental series before and after immune activation. The B cells are derived from inflamed tonsils. The B cells were already activated in vivo in patients, therefore there was no need to stimulate the B cells after isolation. Gene expression profiling was also performed from the same samples. Four B cell subsets: Naïve, germinal center B cells (GC), memory B cells, and plasma cells (PC) were purified by FACS ex vivo from tonsils of 8 human subjects. Methylated CpG island Recovery assay (MIRA) was utilized to enrich for methylated DNA fragments from each sample for microarray analysis. RNA was copurified from the same cell samples for gene expression profiling on microarray. *** This Series reports gene expression profiling data. ***

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The Suppression of Medium Acidosis Improves the Maintenance and Differentiation of Human Pluripotent Stem Cells at High Density in Defined Cell Culture Medium

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