TAMI-16. THREE-DIMENSIONAL ORGANOID CULTURE UNVEILS RESISTANCE TO CLINICAL THERAPIES IN ADULT AND PEDIATRIC GLIOBLASTOMA

2021 ◽  
Vol 23 (Supplement_6) ◽  
pp. vi201-vi201
Author(s):  
Swetha Sundar ◽  
Sajina Shakya ◽  
Lisa Wallace ◽  
Austin Barnett ◽  
Andrew Sloan ◽  
...  
Keyword(s):  
Cellular Proliferation ◽  
Three Dimensional ◽  
Traditional Culture ◽  
Standard Of Care ◽  
Culture Method ◽  
Primary Brain Tumor ◽  
Culture Methods ◽  
Organoid Culture ◽  
Dismal Prognosis ◽  
Organoid Cultures

Abstract BACKGROUND Glioblastoma (GBM) is the most common primary brain tumor with a dismal prognosis. The inherent cellular diversity and interactions within tumor microenvironments represent a significant challenge to effective treatment. Traditional culture methods may mask the complexity of such interactions while three-dimensional (3D) organoid culture systems derived from patient cancer stem cells (CSCs) can preserve cellular complexity and microenvironments. Our objective was to determine whether organoid cultures show increased patterns of resistance to potential clinical therapies compared to traditional sphere cultures. METHODS Adult and pediatric surgical specimens were collected and established as 3D organoids. We created organoid microarrays and visualized bulk and spatially defined differences in cell proliferation using immunohistochemistry (IHC) staining, as well as cell cycle analysis by flow cytometry with 3D regional labeling. We tested the response of CSCs grown in each culture method to temozolomide, ibrutinib, lomustine, ruxolitinib, and radiotherapy using proliferative and viability assays. RESULTS Compared to sphere cultures from the same patient, organoids showed diverse proliferative cell populations and broad resistance to all therapies tested, albeit with both intraspecimen and interspecimen variability in the extent of resistance. Organoid specimens demonstrated a blunt response to current GBM standard of care therapy (combination temozolomide and radiotherapy) and maintained both cellular proliferation in their outer rim and overall structure and viability compared to the matched sphere specimens. CONCLUSIONS Our results suggest that growth of tumor specimens as organoid cultures may better reflect the cellular diversity and clinical reality of GBM therapeutic response. Patient-derived GBM organoids offer a valuable complement to traditional culture methods and may have powerful predictive capability of personalized drug sensitivities and therapeutic resistance.

scholarly journals VIDAS® Enzyme-Linked Immunofluorescent Assay for Detection of Listeria in Foods: Collaborative Study

2000 ◽  
Vol 83 (4) ◽  
pp. 903-918 ◽  
Author(s):  
Vidhya Gangar ◽  
Michael S Curiale ◽  
Armando D’Onorio ◽  
Ann Schultz ◽  
Ronald L Johnson ◽  
...  
Keyword(s):  
False Positive Rate ◽  
Collaborative Study ◽  
False Negative ◽  
Traditional Culture ◽  
Culture Method ◽  
Contamination Level ◽  
Culture Methods ◽  
Positive Rate ◽  
Standard Culture ◽  
Lis Method

Abstract The VIDAS LIS method and the traditional culture methods for detection of Listeria species in food were evaluated in a multilaboratory comparative study. The 6 foods tested were either naturally contaminated or inoculated with 3 different concentrations of Listeria. Results for each food and each contamination level with the VIDAS LIS method were as good as or better than those obtained with the traditional culture method. Of 1558 samples tested, 935 were positive: 839 by the VIDAS method and 809 by standard culture methods. Overall false negative rates were 10.3 and 13.5% for the VIDAS LIS and culture methods, respectively. The false positive rate for the VIDAS LIS assay was 1.4% based on 9 VIDAS LIS positive assays that did not confirm positive by isolation of Listeria. The agreement between the VIDAS LIS and culture methods for all samples tested was 86%.

scholarly journals Comparison of culture methodology for the detection of methicillin-resistant Staphylococcus pseudintermedius in clinical specimens collected from dogs

2017 ◽  
Vol 30 (1) ◽  
pp. 93-98 ◽  
Author(s):  
Matthew E. Saab ◽  
C. Anne Muckle ◽  
Henrik Stryhn ◽  
J. Trenton McClure
Keyword(s):  
High Risk ◽  
Traditional Culture ◽  
Culture Method ◽  
Confirmatory Test ◽  
Clinical Samples ◽  
Culture Methods ◽  
Staphylococcus Pseudintermedius ◽  
Mannitol Salt Agar ◽  
Methicillin Resistant ◽  
Oxacillin Resistance

Methicillin-resistant Staphylococcus pseudintermedius (MRSP) has emerged as a major pathogen in dogs and has been implicated as a hospital-acquired pathogen in veterinary hospitals. We attempted to determine if selective culture methods will detect more MRSP when compared to the traditional culture methods in clinical samples from dogs in Atlantic Canada with a high risk for MRSP infection. Each sample was tested using 4 culture methods: traditional culture; mannitol salt agar with 2 μg/mL of oxacillin (MSAox); enrichment broth (EB) with MSAox; and EB with traditional culture. Detection of penicillin-binding protein 2’, via latex agglutination, was used as a confirmatory test for oxacillin resistance. We analyzed 741 samples from 556 dogs between February 2013 and April 2014. The prevalence of MRSP in samples detected by any method was estimated at 13.4% (95% CI: 11.1–16.0%). When the prevalence of MRSP was determined according to culture method, EB with MSAox detected the highest prevalence (11.2% [9.1–13.7%]), followed by EB with traditional (10.8% [8.8–13.2%]), traditional (10.1% [8.1–12.5%]), and MSAox (8.9% [7.1–11.2%]). The prevalence using the traditional culture method did not differ significantly from any of the 3 selective culture methods. Culture with MSAox detected significantly fewer MRSP than either of the EB methods. The addition of EB to current methodology is recommended, particularly for patients considered at high risk for MRSP infection.

scholarly journals Characterizing Cell Stress and GRP78 in Glioma to Enhance Tumor Treatment

2020 ◽  
Vol 10 ◽  
Author(s):  
Kristie Liu ◽  
Kathleen Tsung ◽  
Frank J. Attenello
Keyword(s):  
Radiation Therapy ◽  
Patient Outcomes ◽  
Cellular Proliferation ◽  
Standard Of Care ◽  
Tumor Grade ◽  
Primary Brain Tumor ◽  
Current Standard ◽  
Unfolded Protein ◽  
The Unfolded Protein Response ◽  
Protein Response

Glioblastoma (GBM) is the most common primary brain tumor, carrying a very poor prognosis, with median overall survival at about 12 to 15 months despite surgical resection, chemotherapy with temozolomide (TMZ), and radiation therapy. GBM recurs in the vast majority of patients, with recurrent tumors commonly displaying increase in resistance to standard of care chemotherapy, TMZ, as well as radiotherapy. One of the most commonly cited mechanisms of chemotherapeutic and radio-resistance occurs via the glucose-regulated protein 78 (GRP78), a well-studied mediator of the unfolded protein response (UPR), that has also demonstrated potential as a biomarker in GBM. Overexpression of GRP78 has been directly correlated with malignant tumor characteristics, including higher tumor grade, cellular proliferation, migration, invasion, poorer responses to TMZ and radiation therapy, and poorer patient outcomes. GRP78 expression is also higher in GBM tumor cells upon recurrence. Meanwhile, knockdown or suppression of GRP78 has been shown to sensitize cells to TMZ and radiation therapy. In light of these findings, various novel developing therapies are targeting GRP78 as monotherapies, combination therapies that enhance the effects of TMZ and radiation therapy, and as treatment delivery modalities. In this review, we delineate the mechanisms by which GRP78 has been noted to specifically modulate glioblastoma behavior and discuss current developing therapies involving GRP78 in GBM. While further research is necessary to translate these developing therapies into clinical settings, GRP78-based therapies hold promise in improving current standard-of-care GBM therapy and may ultimately lead to improved patient outcomes.

scholarly journals Recent Advances in Three-Dimensional Stem Cell Culture Systems and Applications

2021 ◽  
Vol 2021 ◽  
pp. 1-13
Author(s):  
Xiaowen Wu ◽  
Junxiang Su ◽  
Jizhen Wei ◽  
Nan Jiang ◽  
Xuejun Ge
Keyword(s):  
Cell Culture ◽  
Culture System ◽  
Three Dimensional ◽  
Culture Method ◽  
Future Research ◽  
Two Dimensional ◽  
Culture Methods ◽  
Three Dimensional Culture ◽  
Dimensional Cell

Cell culture is one of the most core and fundamental techniques employed in the fields of biology and medicine. At present, although the two-dimensional cell culture method is commonly used in vitro, it is quite different from the cell growth microenvironment in vivo. In recent years, the limitations of two-dimensional culture and the advantages of three-dimensional culture have increasingly attracted more and more attentions. Compared to two-dimensional culture, three-dimensional culture system is better to realistically simulate the local microenvironment of cells, promote the exchange of information among cells and the extracellular matrix (ECM), and retain the original biological characteristics of stem cells. In this review, we first present three-dimensional cell culture methods from two aspects: a scaffold-free culture system and a scaffold-based culture system. The culture method and cell characterizations will be summarized. Then the application of three-dimensional cell culture system is further explored, such as in the fields of drug screening, organoids and assembloids. Finally, the directions for future research of three-dimensional cell culture are stated briefly.

scholarly journals Comparison of Three Nucleic Acid Amplification Tests and Culture for Detection of Group BStreptococcusfrom Enrichment Broth

2019 ◽  
Vol 57 (6) ◽  
Author(s):  
Ji H. Shin ◽  
David T. Pride
Keyword(s):  
Nucleic Acid ◽  
Standard Of Care ◽  
Culture Method ◽  
Nucleic Acid Amplification ◽  
Culture Methods ◽  
Content Type ◽  
Nucleic Acid Amplification Tests ◽  
Hands On ◽  
Group B ◽  
Care Culture

ABSTRACTColonization of the gastrointestinal and genitourinary tracts of pregnant women with group BStreptococcus(GBS) can result in vertical transmission to neonates during labor/delivery. GBS infections in neonates can cause severe complications, such as sepsis, meningitis, and pneumonia. Accurate detection is critical because administration of intrapartum antibiotics can significantly reduce transmission. We compared the clinical sensitivities of three nucleic acid amplification tests (NAATs), the Hologic Panther Fusion GBS, Luminex Aries GBS, and Cepheid Xpert GBS LB assays, to that of the standard of care culture method recommended for GBS screening using 500 vaginal-rectal swab specimens after 18 to 24 h of broth enrichment. We identified 108 positive specimens (21.6%) by culture, while at least 1 of the 3 NAATs was positive for GBS in 155 specimens (31.0%). All 108 specimens positive by culture were also detected by the Panther Fusion assay, while 107/108 (99.1%) were detected by the Cepheid Xpert and Luminex Aries assays. Of the 61 specimens positive by at least 1 NAAT but negative by culture, 24 (39.3%) were positive by all 3 NAATs, suggesting that they represent true positives (TPs). NAATs offer less hands-on time, greater throughput, faster time to result, and potentially greater sensitivity than culture methods, and they should be considered the new gold standard for intrapartum GBS screening.

Automation of Organoid Cultures: Current Protocols and Applications

2021 ◽  
pp. 247255522110245
Author(s):  
Alexandra Louey ◽  
Damián Hernández ◽  
Alice Pébay ◽  
Maciej Daniszewski
Keyword(s):  
Regenerative Medicine ◽  
Drug Screening ◽  
Large Scale ◽  
Three Dimensional ◽  
Hereditary Diseases ◽  
Organoid Culture ◽  
Organoid Cultures

Organoids are three-dimensional, functional structures that mimic in vivo organs. They offer new opportunities for the modeling of cancer and infectious and rare hereditary diseases. Furthermore, the advent of organoid biobanks opens new avenues for drug screening in a personalized fashion and holds much promise for personalized regenerative medicine. Thus, there is a need for reproducible, large-scale organoid generation with minimal variability, making manual approaches impracticable. Here, we review the current use of automation in organoid culture and analysis, using cerebral and retinal organoids as illustrations of current applications. An increased demand for automated organoid platforms is anticipated. Graphical Abstract [Formula: see text]

Deglucohellebrin: A Potent Agent for Glioblastoma Treatment

2020 ◽  
Vol 20 (1) ◽  
pp. 103-110
Author(s):  
Evrysthenis Vartholomatos ◽  
George A. Alexiou ◽  
Georgios S. Markopoulos ◽  
Diamanto Lazari ◽  
Olga Tsiftsoglou ◽  
...  
Keyword(s):  
Anticancer Agents ◽  
Primary Brain Tumor ◽  
Behavioral Alterations ◽  
Endemic Plant ◽  
M Cell ◽  
Apoptotic Pathway ◽  
Cycle Arrest ◽  
Dismal Prognosis ◽  
Mitochondrial Membrane Depolarization ◽  
Induced Changes

Background: Glioblastoma is the most common primary brain tumor in adults with a dismal prognosis. To date, several anticancer agents have been isolated from plants. Helleborus odorus subsp. Cyclophyllus is an endemic plant of the Balcan flora. Herewith, we investigated for the first time, the anti-glioma effect of deglucohellebrin (DGH) extracted from the roots of Helleborus. Methods: We investigated the effect of DGH in U251MG, T98G and U87G glioblastoma cell lines. We selected the T98G cells because of their inherent temozolomide resistance. Results: The IC50 value of reduced viability for DGH was 7x10-5M in U251MG cells, 5x10-5M for the T98G cells and 4x10-5M in U87G cells during 72h treatment. DGH induced G2/M cell cycle arrest, caspace-8 activation and significant mitochondrial membrane depolarization, suggesting the activation of the intrinsic, mitochondrial- dependent apoptotic pathway. DGH and temozolomide induced changes in CDs’ expression in U251MG and T98G cells. In zebrafish, DGH did not induce toxicity or behavioral alterations. Conclusion: The present study is the first to determine the anti-glioma activity of DGH. DGH may be a potent agent for glioblastoma treatment and further studies are needed.

scholarly journals Isolation of Mycobacterium avium Subsp. Paratuberculosis in the Feces and Tissue of Small Ruminants Using a Non-Automated Liquid Culture Method

Animals ◽  
2019 ◽  
Vol 10 (1) ◽  
pp. 20
Author(s):  
Luigi De Grossi ◽  
Davide Santori ◽  
Antonino Barone ◽  
Silvia Abbruzzese ◽  
Matteo Ricchi ◽  
...  
Keyword(s):  
Mycobacterium Avium ◽  
Liquid Culture ◽  
Culture Media ◽  
Small Ruminants ◽  
Culture Method ◽  
Mycobacterium Avium Subsp ◽  
Type I ◽  
Culture Methods ◽  
Specific Pcr ◽  
Mycobacterium Avium Subsp Paratuberculosis

Paratuberculosis is a chronic disease of ruminants caused by Mycobacterium avium subsp. Paratuberculosis (MAP). Since isolation of MAP type I (S) is rarely reported in Italy, our research was aimed at isolating, by an inexpensive liquid culture manual method, this type of MAP isolates. At first, we used an ELISA to point out to serologically positive samples from five flocks. Secondly, we used a fecal direct IS900-qPCR on the ELISA positive samples, in order to detect shedder animals. Feces from IS900-qPCR positive samples were inoculated in solid and liquid culture media. IS900-qPCR was further used to test the growth of MAP isolates in liquid medium, which were further confirmed by f57-qPCR and submitted to typing by specific PCR in order to identify the MAP type. Twenty-eight samples (24 fecal and four tissutal samples) were processed by culture methods, resulting in the isolation of six type I MAP field isolates. Notably, no isolates were recovered by solid media, underlining the utility of this liquid method. Few data about this type of MAP are currently available in Italy, and further analyses should be carried out in order to study the origin and epidemiology of type I strains circulating in Italy.

scholarly journals SEOM clinical guidelines for the treatment of malignant pleural mesothelioma (2020)

2021 ◽  
Vol 23 (5) ◽  
pp. 980-987 ◽  
Author(s):  
E. Nadal ◽  
J. Bosch-Barrera ◽  
S. Cedrés ◽  
J. Coves ◽  
R. García-Campelo ◽  
...  
Keyword(s):  
Survival Benefit ◽  
Early Stage ◽  
Asbestos Exposure ◽  
Standard Of Care ◽  
Tumor Board ◽  
Pleural Mesothelioma ◽  
Therapeutic Approaches ◽  
Dismal Prognosis ◽  
Line Chemotherapy ◽  
Multidisciplinary Tumor Board

AbstractMesothelioma is a rare and aggressive tumour with dismal prognosis arising in the pleura and associated with asbestos exposure. Its incidence is on the rise worldwide. In selected patients with early-stage MPM, a maximal surgical cytoreduction in combination with additional antitumour treatment may be considered in selected patients assessed by a multidisciplinary tumor board. In patients with unresectable or advanced MPM, chemotherapy with platinum plus pemetrexed is the standard of care. Currently, no standard salvage therapy has been approved yet, but second-line chemotherapy with vinorelbine or gemcitabine is commonly used. Novel therapeutic approaches based on dual immunotherapy or chemotherapy plus immunotherapy demonstrated promising survival benefit and will probably be incorporated in the future.

scholarly journals Prospects of immune checkpoint blockade and vaccine-based immunotherapy for glioblastoma

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Stefanie Tietze ◽  
Susanne Michen ◽  
Gabriele Schackert ◽  
Achim Temme
Keyword(s):  
Immune Checkpoint ◽  
Checkpoint Inhibitors ◽  
Primary Brain Tumor ◽  
Immune Checkpoint Blockade ◽  
Checkpoint Blockade ◽  
Oncolytic Viruses ◽  
Therapeutic Vaccines ◽  
Dismal Prognosis ◽  
Tumor Parenchyma ◽  
Immunosuppressive Microenvironment

Abstract Glioblastoma multiforme (GBM) is the most prevalent primary brain tumor endowed with a dismal prognosis. Nowadays, immunotherapy in a particular immune checkpoint blockade and therapeutic vaccines are being extensively pursued. Yet, several characteristics of GBM may impact such immunotherapeutic approaches. This includes tumor heterogeneity, the relatively low mutational load of primary GBM, insufficient delivery of antibodies to tumor parenchyma and the unique immunosuppressive microenvironment of GBM. Moreover, standard treatment of GBM, comprising temozolomide chemotherapy, radiotherapy and in most instances the application of glucocorticoids for management of brain edema, results in a further increased immunosuppression. This review will provide a brief introduction to the principles of vaccine-based immunotherapy and give an overview of the current clinical studies, which employed immune checkpoint inhibitors, oncolytic viruses-based vaccination, cell-based and peptide-based vaccines. Recent experiences as well as the latest developments are reviewed. Overcoming obstacles, which limit the induction and long-term immune response against GBM when using vaccination approaches, are necessary for the implementation of effective immunotherapy of GBM.

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