技术资料

Prenatal cannabis exposure (PCE) influences human brain development,but it is challenging to model PCE using animals and current cell culture techniques. Here,we developed a one-stop microfluidic platform to assemble and culture human cerebral organoids from human embryonic stem cells (hESC) to investigate the effect of PCE on early human brain development. By incorporating perfusable culture chambers,air-liquid interface,and one-stop protocol,this microfluidic platform can simplify the fabrication procedure and produce a large number of organoids (169 organoids per 3.5 cm × 3.5 cm device area) without fusion,as compared with conventional fabrication methods. These one-stop microfluidic assembled cerebral organoids not only recapitulate early human brain structure,biology,and electrophysiology but also have minimal size variation and hypoxia. Under on-chip exposure to the psychoactive cannabinoid,$\Delta$-9-tetrahydrocannabinol (THC),cerebral organoids exhibited reduced neuronal maturation,downregulation of cannabinoid receptor type 1 (CB1) receptors,and impaired neurite outgrowth. Moreover,transient on-chip THC treatment also decreased spontaneous firing in these organoids. This one-stop microfluidic technique enables a simple,scalable,and repeatable organoid culture method that can be used not only for human brain organoids but also for many other human organoids including liver,kidney,retina,and tumor organoids. This technology could be widely used in modeling brain and other organ development,developmental disorders,developmental pharmacology and toxicology,and drug screening. View Publication

Tandemers 2MG,2MG3,3MG and 4MG are derivatives of the potent anti-HIV-1 microbicide candidate griffithsin (GRFT). We compared these compounds anti-HIV-1 activity to GRFT using the viruses CAP206.08 and CAAN5342.A2 that have decreased sensitivity to this lectin. The 2MG and 2MG3 tandemers had similar activity to GRFT against cell-free and cell-associated viruses,while 3MG and 4MG were significantly more potent. Furthermore,the restoration of the 234N or 295N glycan in these viruses,known to increase sensitivity to GRFT,also increased sensitivity to 2MG and 2MG3,and not to 3MG and 4MG. In addition,GRFT resistant viruses generated in-vitro were equally resistant to 2MG and 2MG3 while they had considerably low resistance to 3MG and 4MG. Lastly,all five compounds showed increased inhibitory activity in seminal and vaginal simulants although the effect was more pronounced in the former. These data support further studies of tandemers as potential microbicides. View Publication

Bovine viral diarrhea virus (BVDV) is an important viral disease of cattle that causes immune dysfunction. Macrophages are the key cells for the initiation of the innate immunity and play an important role in viral pathogenesis. In this in vitro study,we studied the effect of the supernatant of BVDV-infected macrophage on immune dysfunction. We infected bovine monocyte-derived macrophages (MDM) with high or low virulence strains of BVDV. The supernatant recovered from BVDV-infected MDM was used to examine the functional activity and surface marker expression of normal macrophages as well as lymphocyte apoptosis. Supernatants from the highly virulent 1373-infected MDM reduced phagocytosis,bactericidal activity and downregulated MHC II and CD14 expression of macrophages. Supernatants from 1373-infected MDM induced apoptosis in MDBK cells,lymphocytes or BL-3 cells. By protein electrophoresis,several protein bands were unique for high-virulence,1373-infected MDM supernatant. There was no significant difference in the apoptosis-related cytokine mRNA (IL-1beta,IL-6 and TNF-a) of infected MDM. These data suggest that BVDV has an indirect negative effect on macrophage functions that is strain-specific. Further studies are required to determine the identity and mechanism of action of these virulence factors present in the supernatant of the infected macrophages. View Publication