The tremendous success of transplantation as a life-saving therapy for end-stage organ failure has increased organ demand. β1,4-N-acetyl-galactosaminyl transferase 2.α(1,3)galactosyl transferase gene knock-out.glycoprotein α(1,3)galactosyl transferase.cytidine monophospho-N-acetylneuraminic acid hydroxylase.This study indicates that OWMs such as cynomolgus monkeys can be used as a relevant model for clinical application of xenotransplantation using TKO pigs. Prolonged CD4 +T cell depletion and low anti-pig antibody titers, which were previously reported important for long-term survival of αGal knock-out (GTKO) xenografts, were not always required for long-term survival of TKO-hTG renal xenografts. Two recipients of TKO-hTG xenografts with low expression of human complement regulatory proteins (CRPs) (TKO-A) survived for 2 and 61 days, whereas six recipients of TKO-hTG xenografts with high CRP expression (TKO-B) survived for 15, 20, 71, 135, 265, and 316 days. Kidney xenografts from TKO-hTG pigs were transplanted into eight cynomolgus recipients without pre-screening for low anti-pig antibody titers. Here, we show that long-term survival of renal xenografts from TKO pigs that express additional human transgenes (hTGs) can be achieved in cynomolgus monkeys. However, previous studies on TKO pigs using old world monkeys (OWMs) have been disappointing because of higher anti-TKO pig antibodies in OWMs than humans. Therefore, it is anticipated that TKO pigs will be better donors for human xenotransplantation. Please contact us with any questions regarding the facility, flow cytometry, or the development of new applications.Porcine cells devoid of three major carbohydrate xenoantigens, αGal, Neu5GC, and SDa (TKO) exhibit markedly reduced binding of human natural antibodies. After hours or weekend access is permitted for trained UConn Health employees. The facility is staffed Monday through Friday from 9 a.m. These include: FlowJo (Treestar Inc.), BD FACS DIVA (BD Bioscience), Cell Quest Pro (BD Bioscience), ModFit LT (Verity), and FCAP Array (BD Bioscience). We also provide several computer workstations with flow cytometry and cytometric bead array analysis software. Additionally, the 15 parameter FACS ARIA II is installed in a Baker Bioprotect III safety cabinet, which facilitates analysis and sorting of potentially biohazardous samples. Our FACS ARIA II cell sorters are both equipped with five lasers one can analyze 15 parameters, and the other is capable of 18 parameter analysis. One of our LSR II instruments is equipped with four lasers and is capable of 13 parameter analysis. Two of our LSR II instruments are equipped with five lasers and are capable of 15 or 18 parameter analysis.
We house two four-color BD FACS Calibur instruments, three BD LSR II instruments, a MACSQuant10, and two BD FACS ARIA II high speed cell sorters. Data should be transferred to this location or to the user's own portable drive after completion of an experiment. The server link is available on the desktop of each computer. Instrument computers can also be used for data analysis, but data acquisition takes precedence over data analysis.Ī centralized flow cytometry server is available for downloading data off of the instruments. As a centralized resource, the flow cytometry facility provides researchers and clinical investigators at UConn Health and surrounding institutions with the ability to perform both multi-parameter analysis and sorting of cell populations into phenotypic, biochemical, and molecular subsets, based on a variety of characteristics including size and shape and fluorescence derived from antibody-tagging or expression of fluorescent molecules.Ĭomputer workstations are available for data analysis.