Health Educ Behav

UL1 TR000433/TR/NCATS NIH HHS/United States5U01CE001957-02/CE/NCIPC CDC HHS/United StatesDA07484/DA/NIDA NIH HHS/United StatesUL1TR000433/TR/NCATS NIH HHS/United StatesR01 DA007484/DA/NIDA NIH HHS/United StatesU01 CE001957/CE/NCIPC CDC HHS/United States2014-03-26T00:00:00Z23863911PMC396656

Fetal Bovine Serum RNA Interferes with the Cell Culture derived Extracellular RNA

Fetal bovine serum (FBS) has been used in eukaryotic cell cultures for decades. However, little attention has been paid to the biological effects associated with RNA content of FBS on cell cultures. Here, using RNA sequencing, we demonstrate that FBS contains a diverse repertoire of protein-coding and regulatory RNA species, including mRNA, miRNA, rRNA, and snoRNA. The majority of them (>70%) are retained even after extended ultracentrifugation in the preparations of vesicle-depleted FBS (vdFBS) commonly utilized in the studies of extracellular vesicles (EV) and intercellular communication. FBS-associated RNA is co-isolated with cell-culture derived extracellular RNA (exRNA) and interferes with the downstream RNA analysis. Many evolutionally conserved FBS-derived RNA species can be falsely annotated as human or mouse transcripts. Notably, specific miRNAs abundant in FBS, such as miR-122, miR-451a and miR-1246, have been previously reported as enriched in cell-culture derived EVs, possibly due to the confounding effect of the FBS. Analysis of publically available exRNA datasets supports the notion of FBS contamination. Furthermore, FBS transcripts can be taken up by cultured cells and affect the results of highly sensitive gene expression profiling technologies. Therefore, precautions for experimental design are warranted to minimize the interference and misinterpretations caused by FBS-derived RNA

EcoSummit 2023 Conference Declaration: Building a Sustainable Wellbeing Future

We - an international community of environmental researchers and practitioners - met in Australia for the 6th International EcoSummit. We acknowledged the traditional custodians of the land on which we met and their elders, past, present, and emerging. We met at a time of multiple, escalating, interconnected crises, including: (1) Biophysical planetary boundaries and being exceeded, including that the climate crisis is here and fast approaching irreversible tipping points, creating a world that human civilization has never experienced. (2) Natural ecosystems and their biodiversity are being lost and degraded resulting in ongoing destruction of sustainable life-supporting functions and services. (3) Social capital is eroding due to runaway inequality and political polarization - people around the world recognize that life is not getting better. (4) Levels of anxiety, depression, and burnout are skyrocketing in parts of the world, while poverty and famine still exist in others. (5) Our economic system remains dependent on fossil fuel, continues to extract renewable natural resources at rates that exceed their regeneration, and has insufficient concern for the loss to biodiversity and damage to valuable ecosystem services, both on land and in the sea. (6) We are faced with increasingly uncontrollable conflicts in various regions linked to resource depletion. We have come together here around the theme of “Building a sustainable and desirable future: Adapting to a changing land and sea-scape.” During this conference, we have presented leading, cutting-edge research showcasing a way forward to achieve sustainable wellbeing for humanity and the rest of nature.</p

Glioblastoma-Associated Microglia Reprogramming Is Mediated by Functional Transfer of Extracellular miR-21

Gliomas are primary, diffusely infiltrating brain tumors. Microglia are innate immune cells in the CNS and make up a substantial portion of the tumor mass. Glioma cells shape their microenvironment, communicating with and reprogramming surrounding cells, resulting in enhanced angiogenesis, immune suppression, and remodeling of the extracellular matrix. Glioma cells communicate with microglia, in part by releasing extracellular vesicles (EVs). Mouse glioma cells stably expressing a palmitoylated GFP to label EVs were implanted intracranially into syngeneic miR-21-null mice. Here, we demonstrate functional delivery of miR-21, regulating specific downstream mRNA targets in microglia after uptake of tumor-derived EVs. These findings attest to EV-dependent microRNA delivery as studied in an in vivo-based model and provide insight into the reprograming of microglial cells by tumor cells to create a favorable microenvironment for cancer progression