Project description:To investigate the changes in liver progenitor cell (LPC) gene expression in response to human amniotic epithelial cell (hAEC) conditioned media. To find supporting evidence to indicate that hAEC conditioned media promotes proliferation and differentiation of LPC’s. To elucidate which pathways are up or down regulated in LPC’s in order to find potential mechanisms and candidate effector factors produced by hAEC’s.
Project description:Myofibroblast activation is a cellular response elicited by a variety of physiological or pathological insults whereby cells initiate a coordinated response intended to eradicate the insult and then revert back to a basal state. However, an underlying theme in various disease states is persistent myofibroblast activation that fails to resolve. Based on multiple observations, we hypothesized that the secreted factors harvested from co-culturing amniotic stem cells might mimic the anti-inflammatory state that cell-free amniotic fluid (AF) elicits. We optimized an amnion epithelial and amniotic fluid cell co-culture system, and tested this hypothesis in the context of myofibroblast activation. However, we discovered that co-cultured amniotic cell conditioned media (coACCM) and AF have opposing effects on myofibroblast activation: coACCM activates the epithelial-mesenchymal transition (EMT) and stimulates gene expression patterns associated with myofibroblast activation, while AF does the opposite. Intriguingly, purified extracellular vesicles (EVs) from AF are necessary and sufficient to activate EMT and inflammatory gene expression patterns, while the EV-depleted AF potently represses these responses. In summary, these data indicate that coACCM stimulates myofibroblast activation, while AF represses it. We interpret these findings to suggest that coACCM, AF, and fractionated AF represent unique biologics that elicit differential cellular responses correlated with a wide variety of pathological states, and therefore could have broad utility in the clinic and the lab.
Project description:This study examines and compares the protein content in conditioned media collected from neural cell types generated from human pluripotent stem cells. Conditioned media was prepared for 48 hours at a final endpoint of differentiation day 12. Both groups are from parental line WTC11 and cultured as a monolayer on matrigel. Both groups contain a transgene cassette for doxycycline-inducible expression of sox9 and nfia. Doxycycline was only included in the iAstro groups, whereas it was omitted in the neural progenitor cell groups.
Project description:Conventional conditions to maintain human embryonic stem cells (hESC) imply the use of inactive mouse embryonic fibroblast (iMEF) as a feeder layer. However, it has suggested that the culture of hESC on iMEF could be an artifact that does not correspond to the in vitro counterpart of the human epiblast. We previously derived and maintained human embryonic stem cells (Amicqui-1 hESC line) on a feeder layer of human amniotic epithelial cells (hAEC). However, the mechanisms involved in the interaction between both cell types to promote the pluripotency still remain unknown. To elucidate if the transcription profile of hESC on hAEC differs from conventional conditions on iMEF, we carried out RNA-seq of Amicqui-1 hESC on both feeder layer conditions (AMIQ hESC-iMEF and AMIQ hESC-hAEC) and on their respective conditioned media (AMIQ hESC-hAEC-CM and AMIQ hESC-iMEF-CM). In this experiment we included H1-hESC-iMEF and hAEC alone.
Project description:Primary human liver sinusoidal endothelial cells from 7 explanted livers were incubated in conditioned media from IMR90 cells undergoing several forms of senescence
Project description:We performed small RNA sequencing of conditioned media collected from breast cancer cells as well as human mammary epithelial cells (HUMEC) in biological replicates.
Project description:Global expression profile of human osteoblast treated with chemotherapy-treated bone marrow stromal cell conditioned media, compared to human osteoblast cells treated with diluent-control bone marrow stromal cell conditioned media. Goal is to identify genes regulated by chemotherapy in osteoblasts.
Project description:Enteroendocrine cells are hormonal secreting cells in the gut. However, as they comprise <1% of the total epithelial cell population, studies on their response to stimuli have been limited. Chang-Graham, Danhof, Engevik, and Tomaro-Duchesneau et al (PMID 31029854) developed a model enteroid system to overcome this limitation. By driving expression of NGN3 in human jejunal enteroids, their system allows for analysis of hormonal secretion and transcriptional analysis in response to a stimulus. To further characterize these NGN3 enteroids, we performed RNASeq on these enteroids in the induced or uninduced state, treated with a media control or Limosilactobacillus reuteri conditioned media. .
