Cytotechnology最新文献

{"title":"Formation of ovarian organoid by co-culture of human endometrial mesenchymal stem cells and mouse oocyte in 3-dimensional culture system","authors":"Mohammad Jafar Bagheri, Mojtaba Rezazadeh Valojerdi, Mojdeh Salehnia","doi":"10.1007/s10616-024-00639-w","DOIUrl":"https://doi.org/10.1007/s10616-024-00639-w","url":null,"abstract":"<p>The purpose of this study was to compare the formation of organoid structures by co-culturing of human endometrial mesenchymal stem cells (hEnMSCs) and mouse germinal vesicle (GV) oocytes in hanging drop and sodium alginate hydrogel co-culture methods. Following the preparation of hEnMSCs and partially denuded mouse germinal vesicle oocytes, they were co-cultured in hanging drop and sodium alginate hydrogel systems as two experimental groups. In respected control groups the hEnMSCs were cultured without oocytes. The organoid formation was evaluated under the inverted microscope in all studied groups during the culture period. The hematoxylin and eosin, alcian blue, periodic acid Schiff, and Masson's trichrome methods, were applied for morphological evaluation and extracellular matrix components staining such as glycosaminoglycan, carbohydrate, and collagen fibers. In addition, the germ cell-like characteristics within the organoid structures were investigated via alkaline phosphatase activity immunocytochemistry for DEAD-box polypeptide 4 (DDX4), and the expression of octamer-binding transcription factor 4 (OCT4), DDX4, and synaptonemal complex protein 3 (SYCP3) genes by real-time RT-PCR. The culturing of hEnMSCs in the hanging drop method led to the formation of organoid structures while this structure was not seen in sodium alginate hydrogel culture. The mean diameter of organoid structures was increased during 4 days of culture in both the experimental and control groups in the hanging drop method, reaching 675.50 ± 18.55 µm and 670.25 ± 21.40 µm, respectively (P &lt; 0.05). Morphological staining indicated some large ovoid cells with euchromatin nuclei in the experimental group, whereas, in the control group cells showed dark and dense nuclei. The extracellular matrix components were deposited in organoid structures in both control and experimental groups. The positive alkaline phosphatase activity and immunocytochemistry for DDX4 confirmed the presence of germ cell-like in the experimental group. Real-time RT-PCR showed a significant increase in the expression of DDX4 and SYCP3 genes and a decrease in the level of OCT4 expression in the experimental group compared with its controls. This study successfully generated organoid structures by co-culture of hEnMSCs and oocytes in the hanging drop method and the hEnMSCs could be differentiated into germ cell-like. This organoid structure has potential applications in regenerative medicine and reproductive biology.</p>","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"174 1","pages":""},"PeriodicalIF":2.2,"publicationDate":"2024-06-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141506711","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Correction: LncRNA-NNT-AS1 contributes to the progression of glioma by miR-582-5p/EZH2 axis","authors":"Tonglin Pan, Min Xue","doi":"10.1007/s10616-024-00636-z","DOIUrl":"https://doi.org/10.1007/s10616-024-00636-z","url":null,"abstract":"","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"34 1","pages":""},"PeriodicalIF":2.2,"publicationDate":"2024-06-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141529576","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Obtusifolin inhibits podocyte apoptosis by inactivating NF-κB signaling in acute kidney injury","authors":"Haiyan Xiang, Yan Wu, Yun Zhang, Yuanhao Hong, Yaling Xu","doi":"10.1007/s10616-024-00638-x","DOIUrl":"https://doi.org/10.1007/s10616-024-00638-x","url":null,"abstract":"<p>Acute kidney injury (AKI) is a common clinical condition and is associated with unacceptable morbidity and mortality. Obtusifolin is an anthraquinone extracted from the seeds of <i>Cassia obtusifolia</i> with anti-inflammatory properties. This study focused on the role and mechanism of obtusifolin in AKI. The mouse podocyte cell line MPC5 was exposed to lipopolysaccharide (LPS) to establish a cell model of AKI. The viability of MPC5 cells treated with obtusifolin and/or LPS was detected by 3-(4, 5-Dimethylthiazol-2-yl)-2,5diphenyltetrazolium bromide assay. Cell apoptosis was analyzed by flow cytometry. The levels of podocyte injury- and apoptosis-related proteins as well as the nuclear factor-kappaB (NF-κB) signaling pathway was examined using western blotting analysis. The renal protective effects of obtusifolin were determined using an LPS-induced mouse model of AKI. Serum creatinine and blood urea nitrogen levels were measured. Hematoxylin–eosin staining of kidney sections was performed to evaluate renal histology. We found that MPC5 cells treated with LPS showed suppressed cell viability (<i>p</i> &lt; 0.01) and increased cell apoptosis (<i>p</i> &lt; 0.001). LPS reduced the protein expression of Bcl-2, nephrin, and synaptopodin as well as increased the protein levels of Bax and Cleaved Caspase-3 in podocytes in a concentration-dependent manner (<i>p</i> &lt; 0.01). In addition, 10 μg/ml LPS-repressed cell viability was rescued by obtusifolin in a concentration-dependent manner (<i>p</i> &lt; 0.01). Moreover, LPS-induced increase in MPC5 cell apoptosis was reversed by obtusifolin treatment (<i>p</i> &lt; 0.01). Obtusifolin administration ameliorated LPS-induced kidney injury and reduced blood urea nitrogen and serum creatinine levels in mice (<i>p</i> &lt; 0.001). Additionally, obtusifolin inhibited LPS-induced activation of NF-κB signaling in vitro and in vivo (<i>p</i> &lt; 0.01). Overall, obtusifolin was effective in protecting renal function against LPS-induced AKI via inactivation of NF-κB signaling, which suggested that obtusifolin may act as a valuable agent for AKI therapy.</p>","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"9 1","pages":""},"PeriodicalIF":2.2,"publicationDate":"2024-06-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141506712","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"From stem cells to extracellular vesicles: a new horizon in tissue engineering and regenerative medicine","authors":"Gajanan Arbade, Jovel Varghese Jose, Arvind Gulbake, Sachin Kadam, Shivaji B. Kashte","doi":"10.1007/s10616-024-00631-4","DOIUrl":"https://doi.org/10.1007/s10616-024-00631-4","url":null,"abstract":"<p>In the fields of tissue engineering and regenerative medicine, extracellular vesicles (EVs) have become viable therapeutic tools. EVs produced from stem cells promote tissue healing by regulating the immune system, enhancing cell proliferation and aiding remodeling processes. Recently, EV has gained significant attention from researchers due to its ability to treat various diseases. Unlike stem cells, stem cell-derived EVs show lower immunogenicity, are less able to overcome biological barriers, and have a higher safety profile. This makes the use of EVs derived from cell-free stem cells a promising alternative to whole-cell therapy. This review focuses on the biogenesis, isolation, and characterization of EVs and highlights their therapeutic potential for bone fracture healing, wound healing, and neuronal tissue repair and treatment of kidney and intestinal diseases. Additionally, this review discusses the potential of EVs for the treatment of cancer, COVID-19, and HIV. In summary, the use of EVs derived from stem cells offers a new horizon for applications in tissue engineering and regenerative medicine.</p>","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"3 1","pages":""},"PeriodicalIF":2.2,"publicationDate":"2024-05-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140940487","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Gellan gum-gelatin based cardiac models support formation of cellular networks and functional cardiomyocytes","authors":"Hanna Vuorenpää, Joona Valtonen, Kirsi Penttinen, Sanna Koskimäki, Emma Hovinen, Antti Ahola, Christine Gering, Jenny Parraga, Minna Kelloniemi, Jari Hyttinen, Minna Kellomäki, Katriina Aalto-Setälä, Susanna Miettinen, Mari Pekkanen-Mattila","doi":"10.1007/s10616-024-00630-5","DOIUrl":"https://doi.org/10.1007/s10616-024-00630-5","url":null,"abstract":"<p>Cardiovascular diseases remain as the most common cause of death worldwide. To reveal the underlying mechanisms in varying cardiovascular diseases, in vitro models with cells and supportive biomaterial can be designed to recapitulate the essential components of human heart. In this study, we analyzed whether 3D co-culture of cardiomyocytes (CM) with vascular network and with adipose tissue-derived mesenchymal stem/stromal cells (ASC) can support CM functionality. CM were cultured with either endothelial cells (EC) and ASC or with only ASC in hydrazide-modified gelatin and oxidized gellan gum hybrid hydrogel to form cardiovascular multiculture and myocardial co-culture, respectively. We studied functional characteristics of CM in two different cellular set-ups and analyzed vascular network formation, cellular morphology and orientation. The results showed that gellan gum-gelatin hydrogel supports formation of two different cellular networks and functional CM. We detected formation of a modest vascular network in cardiovascular multiculture and extensive ASC-derived alpha smooth muscle actin -positive cellular network in multi- and co-culture. iPSC-CM showed elongated morphology, partly aligned orientation with the formed networks and presented normal calcium transients, beating rates, and contraction and relaxation behavior in both setups. These 3D cardiac models provide promising platforms to study (patho) physiological mechanisms of cardiovascular diseases.</p>","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"2011 1","pages":""},"PeriodicalIF":2.2,"publicationDate":"2024-05-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140834239","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"miR-221-3p is upregulated in acute pulmonary embolism complicated with pulmonary hypertension and promotes pulmonary arterial smooth muscle cells proliferation and migration by inhibiting PTEN","authors":"Lei Tang, Shuai Niu, Jinwei Xu, Wei Lu, Li Zhou","doi":"10.1007/s10616-024-00628-z","DOIUrl":"https://doi.org/10.1007/s10616-024-00628-z","url":null,"abstract":"<p>Pulmonary arterial smooth muscle cells (PASMCs) functions are associated with the pathogenesis of pulmonary hypertension (PH) which is a life-threatening complication of acute pulmonary embolism (APE). This study sought to explore the expression pattern of microRNA (miR)-221-3p in APE-PH patients and its role in PASMCs proliferation and migration. The clinical data and venous blood of APE-PH patients were collected. The expression levels of miR-221-3p and phosphatase and tensin homolog (PTEN) in serum were determined, followed by receiver operator characteristic curve analysis of miR-221-3p diagnostic efficacy. PASMCs were transfected with miR-221-3p mimics and PTEN-overexpressed vector, followed by assessment of cell viability, proliferation, and migration through cell counting kit-8, 5‐ethynyl‐2′‐deoxyuridine, Transwell, and wound healing assays. The binding between miR-221-3p and PTEN 3′UTR region was testified by the dual-luciferase assay. miR-221 was upregulated in the serum of APE-PH patients and presented with good diagnostic efficacy with 1.155 cutoff value, 66.25% sensitivity, and 67.50% specificity. miR-221 was negatively correlated with PTEN in APE-PH patients. miR-221 overexpression facilitated PASMCs proliferation and migration in vitro. miR-221-3p bound to PTEN 3′UTR region to decrease PTEN protein levels. PTEN overexpression abolished the promotive role of miR-221-3p in PASMCs. Overall, miR-221-3p targeted PTEN to facilitate PASMC proliferation and migration.</p>","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"55 1","pages":""},"PeriodicalIF":2.2,"publicationDate":"2024-04-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140834109","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"PABPC1 silencing inhibits pancreatic cancer cell proliferation and EMT, and induces apoptosis via PI3K/AKT pathway","authors":"Changren Zhu, Cuimei Wang, Xiaodong Wang, Shuangshuang Dong, Qing Xu, Jun Zheng","doi":"10.1007/s10616-024-00626-1","DOIUrl":"https://doi.org/10.1007/s10616-024-00626-1","url":null,"abstract":"<p>Pancreatic cancer is difficult to manage owing to the challenges involved in its treatment and nursing. This study aimed to clarify the roles and mechanisms of action of Poly (A)-binding protein cytoplasmic 1 (PABPC1) on pancreatic cancer. The expression of PABPC1 in pancreatic cancer tissues and cell lines was detected using RT-qPCR and western blotting. The effects of PABPC1 on proliferation, apoptosis, epithelial-mesenchymal transition (EMT), and the PI3K/AKT signaling pathway in pancreatic cancer cells were further investigated using MTT assays, flow cytometry, and western blotting. The expression of PABPC1 was significantly upregulated in pancreatic cancer tissues and cells, whereas PABPC1 downregulation inhibited pancreatic cancer cell proliferation, induced apoptosis, decreased the expression of EMT-associated proteins, and exerted a regulatory effect by inhibiting the PI3K/AKT signaling pathway. In addition, the findings indicated that PABPC1 over-expression significantly promoted pancreatic cancer cell proliferation, inhibited apoptosis, decreased the expression of E-cadherin, enhanced N-cadherin expression, and activating the PI3K/AKT signaling pathway. PABPC1 silencing significantly inhibited proliferation and EMT and induced apoptosis in pancreatic cancer cells. These findings provide novel insights into the role of PABPC1 in the development of pancreatic cancer.</p>","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"94 1","pages":""},"PeriodicalIF":2.2,"publicationDate":"2024-04-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140635551","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Pyrroloquinoline quinone protects against murine hepatitis virus strain 3-induced fulminant hepatitis by inhibiting the Keap1/Nrf2 signaling","authors":"Zunguo Pu, Fei Ge, Yaqing Zhou, Aiming Liu, Chao Yang","doi":"10.1007/s10616-024-00627-0","DOIUrl":"https://doi.org/10.1007/s10616-024-00627-0","url":null,"abstract":"<p>Fulminant hepatitis (FH) is a life-threatening clinical liver syndrome characterized by substantial hepatocyte necrosis and severe liver damage. FH is typically associated with severe oxidative stress, inflammation, and mitochondrial dysfunction. Pyrroloquinoline quinone (PQQ), a naturally occurring redox cofactor, functions as an essential nutrient and antioxidant and reportedly inhibits oxidative stress and exerts potent anti-inflammatory effects. In the present study, we aimed to evaluate the therapeutic efficacy of PQQ in murine hepatitis virus strain 3 (MHV-3)-induced FH and examined the underlying mechanism. An MHV-3-induced FH mouse model was established for in vivo examination<i>.</i> Liver sinusoidal endothelial cells (LSECs) were used for in vitro experiments. Herein, we observed that PQQ supplementation significantly attenuated MHV-3-induced hepatic injury by suppressing inflammatory responses and reducing oxidative stress. Mechanistically, PQQ supplementation ameliorated MHV-3-induced hepatic damage by down-regulating the Keap1/Nrf2 signaling pathway in vivo and in vitro. Furthermore, Nrf2 small interfering RNA targeting LSECs abrogated the PQQ-mediated protective effects against MHV-3-related liver injury. Our results deepen our understanding of the hepatoprotective function of PQQ against MHV-3-induced liver injury and provide evidence that alleviating oxidative stress might afford a novel therapeutic strategy for treating FH.</p>","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"39 1","pages":""},"PeriodicalIF":2.2,"publicationDate":"2024-04-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140608961","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Stable two- and three-dimensional cholangiocyte culture systems from extrahepatic bile ducts of biliary atresia patients: use of structural and functional bile duct epithelium models for in vitro analyses","authors":"Ai Shimamura, Mayumi Higashi, Kazuya Nagayabu, Shigeru Ono","doi":"10.1007/s10616-024-00620-7","DOIUrl":"https://doi.org/10.1007/s10616-024-00620-7","url":null,"abstract":"<p>We herein report two- (2D) and three-dimensional (3D) culture methods of cholangiocytes originating from extrahepatic bile ducts of biliary atresia (BA) patients. Cells were stabilized for in vitro analyses, and 3D culture by two different methods showed the structural and functional features of cholangiocytes in the gel scaffold. First, cells were obtained from gallbladder contents or resected tissues of patients at surgery and then cultured in our original conditioned medium with a cocktail of signaling inhibitors that maintains the immaturity and amplification of cells. Cells were immortalized by inducing SV40T and hTERT genes using lentivirus systems. Immunostaining with CK19 and Sox9 antibodies confirmed the cells as cholangiocytes. 3D organoids were formed in Matrigel in two different ways: by forming spheroids or via vertical growth from 2D cell sheets (2 + 1D culture). Organoids generated with both methods showed the uptake and excretion of rhodamine-123, and duct-like structures were also found. Our culture methods are simpler than previously reported methods and still show the structural and functional characteristics of cholangiocytes. Thus, this system is expected to be useful for the in vitro investigation of cholangiocyte damage or regeneration in BA patients.</p>","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"43 1","pages":""},"PeriodicalIF":2.2,"publicationDate":"2024-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140564879","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The effect of TFAP2A/ANXA8 axis on ferroptosis of cervical squamous cell carcinoma (CESC) in vitro","authors":"Yuehua Sheng, Huiqing Ding, Jiaqing Zhou, Yuejing Wu, Kejun Xu, Fan Yang, Yongming Du","doi":"10.1007/s10616-024-00619-0","DOIUrl":"https://doi.org/10.1007/s10616-024-00619-0","url":null,"abstract":"<p>Potential role and associated mechanisms of Annexin A8 (ANXA8), a member of the Annexins family, in cervical squamous cell carcinoma (CESC) are still unclear, despite being upregulated in various malignant tumors. Here, we observed a notably elevated expression of ANXA8 in CESC cells. The inhibition of ANXA8 amplified the susceptibility of CESC cells to Erastin and sorafenib-induced ferroptosis, whereas it exerted minimal influence on DPI7 and DPI10-induced ferroptosis. The results from the Fe²⁺ concentration assay showed no significant correlation between <i>ANXA8</i> gene knockdown and intracellular Fe²⁺ concentration induced by ferroptosis inducers. Western blot analysis demonstrated that the knockdown of <i>ANXA8</i> did not alter ACSL4 and LPCAT levels under ferroptosis-inducing conditions, but it did result in a reduction in intracellular GSH levels induced by the ferroptosis inducer. Subsequently, we identified TFAP2A as an upstream transcription factor of <i>ANXA8</i>, which plays a role in regulating cell ferroptosis. The knockdown of <i>TFAP2A</i> significantly elevated MDA levels and depressed GSH levels in the presence of a ferroptosis inducer, thereby inhibiting cell ferroptosis. However, this inhibitory effect could be reversed by <i>ANXA8</i> overexpression. Therefore, our research suggests that the TFAP2A/ANXA8 axis exerts regulatory control over ferroptosis in CESC cells by mediating GSH synthesis in System Xc.</p>","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"49 1","pages":""},"PeriodicalIF":2.2,"publicationDate":"2024-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140564537","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}