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High-quality single-cell transcriptomics from ovarian histological sections during folliculogenesis
Get PDFHigh-quality, straightforward single-cell RNA sequencing (RNA-seq) with spatial resolution remains challenging. Here, we developed DRaqL (direct RNA recovery and quenching for laser capture microdissection), an experimental approach for efficient cell lysis of tissue sections, directly applicable to cDNA amplification. Single-cell RNA-seq combined with DRaqL allowed transcriptomic profiling from alcohol-fixed sections with efficiency comparable with that of profiling from freshly dissociated cells, together with effective exon-exon junction profiling. The combination of DRaqL with protease treatment enabled robust and efficient single-cell transcriptome analysis from formalin-fixed tissue sections. Applying this method to mouse ovarian sections, we were able to predict the transcriptome of oocytes by their size and identified an anomaly in the size-transcriptome relationship relevant to growth retardation of oocytes, in addition to detecting oocyte-specific splice isoforms. Furthermore, we identified differentially expressed genes in granulosa cells in association with their proximity to the oocytes, suggesting distinct epigenetic regulations and cell-cycle activities governing the germ-soma relationship. Thus, DRaqL is a versatile, efficient approach for high-quality single-cell RNA-seq from tissue sections, thereby revealing histological heterogeneity in folliculogenic transcriptome
Current Status and Future Challenges for Continuous Positive Airway Pressure Adherence for Obstructive Sleep Apnea : A Narrative Review
Get PDF持続陽庄呼吸療法(continuous positive airway pressure: CPAP)は中等症以上の閉塞性睡眠時無呼吸(obstructive sleep apnea: OSA)に対する標準的治療法である。CPAPは睡眠する際に鼻にマスクを装着して陽圧の空気を気道に送気することによって上気道が虚脱するのを防ぐ装置である。したがって、CPAPを装着していないときは、上気道の開存性を支える圧のスプリントが無いため、上気道が虚脱してOSAを引き起こす。つまり、CPAPはOSAを根治させる治療機器ではなく矯正機器であると言える。よって患者はCPAPを睡眠する際には必ず装着する必要があり、その使用状況が治療効果にも反映される。CPAP使用状況を意味するCPAPアドヒアランスは現実的には満足できるレベルにはない。CPAPアドヒアランスに影響を与える要因は、適切なマスク選択、患者が問題を抱えた際のトラブルシューティングにおける医療者側の技量、CPAP機器の設定調整、OSAの疾患多様性、さらには患者の睡眠衛生など多岐にわたる。これらの問題解決は医師のみでは不可能であり、看護師、臨床検査技師、公認心理師など多職種が連携して様々な側面からCPAPアドヒアランス向上に取り組むべきであると考える
内耳前庭細胞由来因子とオルガノイド培養を組み合わせたES細胞から内耳前庭有毛細胞への分化誘導
Get PDFVestibular hair cells (V-HCs) residing in the inner ear have important roles related to balance. Although differentiation of pluripotent stem cells into HCs has been shown, an effective method has yet to be established. We previously reported that use of vestibular cell-derived conditioned medium (V-CM) was helpful to induce embryonic stem (ES) cells to differentiate into V-HC-like cells in two-dimensional (2D) cultures of ES-derived embryoid bodies (EBs). In the present report, V-CM was used with three-dimensional (3D) cultures of EBs, which resulted in augmented expression of V-HC-related markers (Math1, Myosin6, Brn3c, Dnah5), but not of the cochlear HC-related marker Lmod3. Gene expression analyses of both 2D and 3D EBs cultured for two weeks revealed a greater level of augmented induction of HC-related markers in the 3D-cultured EBs. These results indicate that a 3D culture in combination with use of V-CM is an effective method for producing V-HCs.博士（医学）・甲第877号・令和5年3月15
複数異種移植マウスモデルにおけるPentagamavunone-1の膵臓癌に対する単剤および併用療法としての前臨床評価
Get PDFWe previously reported that pentagamavunone-1 (PGV-1) effectively inhibited cell proliferation in many types of human tumors, including pancreatic cancer, by inducing M phase (prometaphase) arrest, senescence, and apoptosis with few side effects. However, a detailed evaluation of the effects of PGV-1 on pancreatic cancer cells in an in vivo setting has not yet been conducted. The present study investigated the potential efficacy of PGV-1 as both monotherapy and combination therapy for pancreatic cancer using multiple xenograft mouse assays. A cell-line derived xenograft model (CDX-M) with pancreatic cancer cell line and a patient-derived xenograft mouse model (PDX-M) using resected pancreatic cancer samples without neoadjuvant chemotherapy were established in both heterotopic and orthotopic manners. PGV-1 effectively suppressed tumor formation at the heterotopic and orthotopic sites in CDX-M than in untreated mice. Combination therapy with PGV-1 and gemcitabine more effectively suppressed tumor formation than monotherapy with PGV-1 or gemcitabine when administered after tumor formation. Monotherapy with PGV-1 or gemcitabine less effectively suppressed tumor formation in PDX-M than in CDX-M, whereas combination therapy with PGV-1 and gemcitabine more effectively suppressed tumor formation. PGV-1 as monotherapy and combination therapy with gemcitabine effectively inhibited tumor formation and has potential as an anticancer candidate for pancreatic cancer.権利情報：© The Author(s) 2022. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/
