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Cryopreservation and recovery of human endometrial epithelial cells with high viability, purity, and functional fidelity
© 2016 American Society for Reproductive Medicine Objective To develop a protocol for cryopreservation and recovery of human endometrial epithelial cells (eECs) retaining molecular and functional characteristics of endometrial epithelium in vivo. Design In vitro study using human endometrial cells. Setting University research laboratory. Patient(s) Endometrial biopsies were obtained from premenopausal women undergoing benign gynecologic procedures. Intervention(s) Primary eECs were cryopreserved in 1% fetal bovine serum/10% dimethylsulfoxide in Defined Keratinocyte Serum-Free Medium (KSFM). Recovered cells were observed for endometrial stromal fibroblast (eSF) contamination and subsequently evaluated for morphology, gene expression, and functional characteristics of freshly cultured eECs and in vivo endometrial epithelium. Main Outcome Measure(s) Analysis of eEC morphology and the absence of eSF contamination; evaluation of epithelial-specific gene and protein expression; assessment of epithelial polarity. Result(s) Endometrial epithelial cells recovered after cryopreservation (n = 5) displayed epithelial morphology and expressed E-cadherin (CDH1), occludin (OCLN), claudin1 (CLDN1), and keratin18 (KRT18). Compared with eSF, recovered eECs displayed increased (P<.05) expression of epithelial-specific genes AREG, CDH1, DEFB4A, MMP7, and WNT7A, while exhibiting low-to-undetectable (P<.05) stromal-specific genes COL6A3, HOXA11, MMP2, PDGFRB, and WNT5A. Recovered eECs secreted levels of cytokines and growth factors similarly to freshly cultured eECs. Recovered eECs could form a polarized monolayer with high transepithelial electrical resistance (TER) and impermeability to small molecules, and expressed apical/basolateral localization of CDH1 and apical localization of OCLN. Conclusion(s) We have developed a protocol for cryopreservation of eECs in which recovered cells after thawing demonstrate morphologic, transcriptomic, and functional characteristics of human endometrial epithelium in vivo.
© 2015 IEEE. Heterogeneous networks, HetNets, are known to enhance the bandwidth efficiency and throughput of wireless networks by more effectively utilizing the network resources. However, the higher density of users and access points in HetNets introduces significant inter-user interference that needs to be mitigated through complex and sophisticated interference cancellation schemes. Moreover, due to significant channel attenuation and the presence of hardware impairments, e.g. phase noise and amplifier nonlinearities, the vast bandwidth in the millimeterwave band has not been fully utilized to date. In order to enable the development of multi-Gigabit per second wireless networks, we introduce a novel millimeter-wave HetNet paradigm, termed hybrid HetNet, which exploits the vast bandwidth and propagation characteristics in the 60 GHz and 70-80 GHz bands to reduce the impact of interference in HetNets. Simulation results are presented to illustrate the performance advantage of hybrid HetNets with respect to traditional networks. Next, two specific transceiver structures that enable hand-offs from the 60 GHz band, i.e. the V-band to the 70-80 GHz band, i.e. the E-band, and vice versa are proposed. Finally, the practical and regulatory challenges for establishing a hybrid HetNet are outlined.