BEGIN:VCALENDAR VERSION:2.0 CALSCALE:GREGORIAN PRODID:iCalendar-Ruby BEGIN:VEVENT CATEGORIES:Seminar/Colloquia DESCRIPTION:Presenting on “Cells in Machines and Things we Discovered Along the Way”\n\nThomas Boland\, PhD\, professor of metallurgical\, materials a nd biomedical engineering and director of biomedical engineering program at The University of Texas at El Paso\, will speak on Thursday\, March 2\, 20 23 at 10:00 am CST in Whitaker 218.\n\nAbstract: When we first described in kjet bioprinting (TIB) of cells in 2003\, the devices were treated like a b lack-box. Quantifying the input and measuring the output has become the ha llmark of not only our group but\, indeed\, the entire field of bioprinting . Most of this is understandable\, as it is virtually impossible to charact erize cells that are inside the printing orifices\, extrusion needles\, or drops ejected from ribbons. Also understandable is then that we may be miss ing out on important information about cell biology\, for example\, what ma y be happening inside the printer or shortly after the bioprinting event. W e will describe here some recent discoveries of cell behavior due to inkjet bioprinting. \n\nFor these experiments\, we use primary adult dermal fibro blasts which were expanded for 2-3 passages upon receiving. The cells were harvested and resuspended in PBS and bioprinted into a 96-well plate with p luriSTEM media. Cells were then transferred either into precoated 96-well p lates or 20 µL drops were pipetted for hanging drop culture using a multich annel pipettor. IPC to differentiation protocols was applied and the induct ion was begun approximately 45 mins after TIB for the cells that were trans ferred to the plate immediately after printing. When differentiating aggreg ates\, the initiation happened 45 mins after the aggregates were transferre d into the 96 wells. Preliminary results indicate that all cells expressed the 3 pluripotency markers oct-4\, nanog\, and sox-2. After differentiation protocols\, the cells stained positively for tropinin-3 for the cardiomyo cyte differentiation protocol. The cells also elongated and became more ca rdiomyocyte-like in their morphology. We analyzed bulk RNA seq data and ou r preliminary results showed upregulation of some genes that have been impl icated as stem cell markers: EPCAM\, LEFTY1\, ZFP42\, and TEX19. In additio n\, differential expression of genes associated with pluripotency-relevant pathways show some pathways are off like the MAPK/p38\, MAPK/JNK1-3 which w ould be expected for a pluripotent state\, but at this time the activation of the hippo pathway is inconclusive with TAZ highly upregulated but YAP sl ightly downregulated. Similarly\, GSK3B is off and TGFB1\, LIF/PIK3\, and A KT1 are on as expected for pluripotency\, but SMAD2/3 and CTNNB1. Examining the gene network of upregulated genes\, one can clearly distinguish the pi votal role of FOS\, FOXO1\, and PIK3 all related to pluripotency. In conclu sion\, bioprinted fibroblasts will at least temporarily adopt a more primit ive or dedifferentiated state\, reminiscent of pluripotency. While immunoch emistry shows the classic transcription factors required for pluripotency\, gene expression shows a more nuanced picture of the transformations that o ccur upon printing. Understanding these transformations\, even if temporary will be crucial when trying to build tissues using bioprinting technologie s.\n\nRegistration is required to attend virtually. Please register. DTEND:20230302T170000Z DTSTAMP:20260314T015207Z DTSTART:20230302T160000Z GEO:38.649125;-90.303135 LOCATION:Uncas A. Whitaker Hall\, 218 SEQUENCE:0 SUMMARY:BME Seminar: Thomas Boland\, PhD UID:tag:localist.com\,2008:EventInstance_42092511895082 URL:https://happenings.washu.edu/event/bme_seminar_thomas_boland_phd END:VEVENT END:VCALENDAR