Organoids Pub Date : 2024-07-05 DOI: 10.3390/organoids3030010

M. N. H. Tang, M. Moosajee, Najam A Sharif, G. A. Limb, K. Eastlake

{"title":"Heparin-Binding Epidermal-like Growth Factor (HB-EGF) Reduces Cell Death in an Organoid Model of Retinal Damage","authors":"M. N. H. Tang, M. Moosajee, Najam A Sharif, G. A. Limb, K. Eastlake","doi":"10.3390/organoids3030010","DOIUrl":"https://doi.org/10.3390/organoids3030010","url":null,"abstract":"In zebrafish and various mammalian species, HB-EGF has been shown to promote Müller glia proliferation and activation of repair mechanisms that have not been fully investigated in human retina. In the current study, 70- to 90-day-old human retinal organoids were treated with 20 μM 4-hydroxytamoxifen (4-OHT), and CRX, REC, NRL, PAX6, VIM, GFAP, and VSX2 gene and protein expression were assessed at various times points after treatment. Organoids with or without 4-OHT-induced damage were then cultured with HB-EGF for 7 days. We showed that 20 μM 4-OHT caused a reduction in the number of recoverin-positive cells; an increase in the number of TUNEL-positive cells; and downregulation of the photoreceptor gene markers CRX, NRL, and REC. Culture of organoids with HB-EGF for 7 days after 4-OHT-induced damage caused a marked reduction in the number of TUNEL-positive cells and small increases in the number of Ki67-positive cells and PAX6 and NOTCH1 gene expression. The current results suggest that treatment of human ESC-derived retinal organoids with 4-OHT may be used as a model of retinal degeneration in vitro. Furthermore, HB-EGF treatment of human retinal organoids increases proliferating Müller cells, but only after 4-OHT induced damage, and may be an indication of Muller reactivity in response to photoreceptor damage. Further studies will aim to identify factors that may induce Müller cell-mediated regeneration of the human retina, aiding in the development of therapies for retinal degeneration.","PeriodicalId":513546,"journal":{"name":"Organoids","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-07-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141673955","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Development and Optimization of a Lactate Dehydrogenase Assay Adapted to 3D Cell Cultures 开发并优化适用于三维细胞培养的乳酸脱氢酶检测方法

Organoids Pub Date : 2024-06-05 DOI: 10.3390/organoids3020008

Héloïse Castiglione, Lucie Madrange, Thomas Lemonnier, Jean-Philippe Deslys, Frank Yates, Pierre-Antoine Vigneron

{"title":"Development and Optimization of a Lactate Dehydrogenase Assay Adapted to 3D Cell Cultures","authors":"Héloïse Castiglione, Lucie Madrange, Thomas Lemonnier, Jean-Philippe Deslys, Frank Yates, Pierre-Antoine Vigneron","doi":"10.3390/organoids3020008","DOIUrl":"https://doi.org/10.3390/organoids3020008","url":null,"abstract":"In recent years, 3D cell culture systems have emerged as sophisticated in vitro models, providing valuable insights into human physiology and diseases. The transition from traditional 2D to advanced 3D cultures has introduced novel obstacles, complicating the characterization and analysis of these models. While the lactate dehydrogenase (LDH) activity assay has long been a standard readout for viability and cytotoxicity assessments in 2D cultures, its applicability in long-term 3D cultures is hindered by inappropriate normalization and low LDH stability over time. In response to these challenges, we propose an optimization of LDH assays, including a crucial normalization step based on total protein quantification and a storage method using an LDH preservation buffer. We applied it to compare unexposed cerebral organoids with organoids exposed to a toxic dose of valproic acid, and showed efficient normalization of cellular viability as well as enhanced LDH stability within the buffer. Importantly, normalized LDH activity results obtained were independent of organoid dimension and cell density. This refined LDH assay, tailored to address 3D culture constraints, allows for the transposition of this routine test from 2D to 3D cultures.","PeriodicalId":513546,"journal":{"name":"Organoids","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-06-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141383713","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Treatment of Canine Type 1 Diabetes Mellitus: The Long Road from Twice Daily Insulin Injection towards Long-Lasting Cell-Based Therapy 犬 1 型糖尿病的治疗：从每日两次注射胰岛素到长效细胞疗法的漫长之路

Organoids Pub Date : 2024-04-04 DOI: 10.3390/organoids3020006

Flavia C. M. Oliveira, A. Voorbij, Elisa C. Pereira, Leonor M. M. Alves e Almeida, Geanne R. Moraes, Joana T. De Oliveira, Boyd H. T. Gouw, Sabrina A. M. Legatti, Hans S. Kooistra, B. Spee, Andre M. C. Meneses, Louis C. Penning

{"title":"Treatment of Canine Type 1 Diabetes Mellitus: The Long Road from Twice Daily Insulin Injection towards Long-Lasting Cell-Based Therapy","authors":"Flavia C. M. Oliveira, A. Voorbij, Elisa C. Pereira, Leonor M. M. Alves e Almeida, Geanne R. Moraes, Joana T. De Oliveira, Boyd H. T. Gouw, Sabrina A. M. Legatti, Hans S. Kooistra, B. Spee, Andre M. C. Meneses, Louis C. Penning","doi":"10.3390/organoids3020006","DOIUrl":"https://doi.org/10.3390/organoids3020006","url":null,"abstract":"For over 150 years, researchers have studied the (patho)physiology of the endocrine pancreas and devised treatment options for diabetes mellitus (DM). However, no cure has been developed so far. In dogs, diabetes mellitus type 1 (T1DM) is the most common presentation. Treatment consists of twice daily insulin injections, monitored by spatial blood glucose measurements. Even though dogs were instrumental in the discovery of insulin and islet transplantations, the treatment in diabetic dogs has remained unchanged for decades. Providing twice daily insulin injections is demanding for both owners and dogs and may result in hypoglycaemic events, creating the need for new treatment strategies. Novel regenerative medicine-based tools, such as improved β-cell culture protocols and artificial devices, have sparked hope for a cure. In human medicine, emerging technologies such as the transplantation of insulin-producing β-cells, generated by stem cell differentiation, with or without an encapsulation device, are currently tested in phase I/II clinical trials. As the pathogenesis of T1DM is remarkably similar between humans and dogs, novel treatment methods could be implemented in canine medicine. This review briefly summarises the physiology of the canine endocrine pancreas and the pathophysiology of canine DM before exploring current and possible future treatment options for canine DM.","PeriodicalId":513546,"journal":{"name":"Organoids","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-04-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140742350","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Generation of Trophoblast Organoids from Chorionic Villus Sampling 从绒毛取样中生成滋养层细胞器质

Organoids Pub Date : 2024-03-05 DOI: 10.3390/organoids3010005

B. V. van Rijn, Diane Van Opstal, Nicole van Koetsveld, Maarten Knapen, Joost Gribnau, Olivier Schäffers

引用次数: 0

Visualization of Vascular Perfusion of Human Pancreatic Cancer Tissue in the CAM Model and Its Impact on Future Personalized Drug Testing CAM 模型中人类胰腺癌组织血管灌注可视化及其对未来个性化药物测试的影响

Organoids Pub Date : 2024-01-08 DOI: 10.3390/organoids3010001

Andreas Ettner-Sitter, Agata Montagner, Jonas Kuenzel, Kathrin Brackmann, Maximilian Schäfer, Robert Schober, Florian Weber, Thiha Aung, Christina Hackl, Silke Haerteis

{"title":"Visualization of Vascular Perfusion of Human Pancreatic Cancer Tissue in the CAM Model and Its Impact on Future Personalized Drug Testing","authors":"Andreas Ettner-Sitter, Agata Montagner, Jonas Kuenzel, Kathrin Brackmann, Maximilian Schäfer, Robert Schober, Florian Weber, Thiha Aung, Christina Hackl, Silke Haerteis","doi":"10.3390/organoids3010001","DOIUrl":"https://doi.org/10.3390/organoids3010001","url":null,"abstract":"Although significant improvements have been made in the treatment of pancreatic cancer, its prognosis remains poor with an overall 5-year survival rate of less than 10%. New experimental approaches are necessary to develop novel therapeutics. In this study, the investigation of pancreatic cancer tissue growth in the chorioallantoic membrane (CAM) model and the subsequent use of indocyanine green (ICG) injections for the verification of intratumoral perfusion was conducted. ICG was injected into the CAM vasculature to visualize the perfusion of the tumor tissue. The presence of metastasis was investigated through PCR for the human-specific ALU element in the liver of the chicken embryo. Additionally, the usage of cryopreserved pancreatic tumors was established. Intratumoral perfusion of tumor tissue on the CAM was observed in recently obtained and cryopreserved tumors. ALU-PCR detected metastasis in the chick embryos’ livers. After cryopreservation, the tissue was still vital, and the xenografts generated from these tumors resembled the histological features of the primary tumor. This methodology represents the proof of principle for intravenous drug testing of pancreatic cancer in the CAM model. The cryopreserved tumors can be used for testing novel therapeutics and can be integrated into the molecular tumor board, facilitating personalized tumor treatment.","PeriodicalId":513546,"journal":{"name":"Organoids","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139446044","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

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