organoid cultures derived from patients

LHJ Using a 3D organoid system, we report success in long-term culture of prostate cancer from biopsy specimens and circu D, Heat map displaying the mutational signature contributions of PTC tumors and the paired organoids. According to best practice guidelines, sequence data were mapped to the human reference genome (hg19) using Burrows-Wheeler alignment with maximal exact matches (BWA-MEM) version 0.7.12 followed by marked duplicates, merging of lanes, and realignment of small insertions and deletions (indels). 2023 Jun 13;18(6):1255-1270. doi: 10.1016/j.stemcr.2023.05.009. Patient-derived tumor organoid-based high-throughput screening platforms are also widely used for discovering anti-cancer drugs. 3 [20]). Would you like email updates of new search results? Patientderived organoids (PDOs) represent promising preclinical models in various tumor types. Bars represent meansSEM of 3 independent experiments. Putative targets of the tested anticancer drugs are listed on the left. 81772737), Shenzhen Key Basic Research Program (grant No. Each data point represents the meanSEM of 3 technical duplicates. 2021 Feb 18;12(1):1117. doi: 10.1038/s41467-021-21300-6. However, many drawbacks hamper these 2 models for clinical use. 4A, 5A, and 5B). Then, culture as patient-derived organoids (PDOs) by the hanging drop method. Front Cell Dev Biol. We aimed to understand the mechanisms of resistance to chemoradiotherapy in rectal cancer using patient derived organoid models . These deparaffinized tissues and organoids sections were then subjected to H&E and immunohistochemistry staining. Search for other works by this author on: Department of Breast and Thyroid Surgery, Shenzhen Second Peoples Hospital, Department of Nuclear Medicine, Peking University Shenzhen Hospital. These reagents were carefully diluted with dimethyl sulfoxide (DMSO) according to the respective manufacturers instructions to prepare stock solutions, and stored at 80 C. Cancer immunotherapies immune checkpoint inhibitors as patient-derived organoids are under investigation as a personalized platform for drug discovery and for predicting clinical efficacy of chemotherapies and targeted agents. On arrival, the fresh specimens were washed and split into several smaller pieces. , Cesari V, Visani M, et al. 5) (20). Patient-derived breast tumor organoids developed in our laboratory consist of a mix of multiple tumor-derived cell populations, and thus represent a better approximation of tumor cell diversity and milieu than the established 2D cancer cell lines. Copy number landscape of patient derived prostate cancer organoid lines, Figure 2. Genomic DNA from blood (n=8), PTC tissues (n=8), and tumor-derived organoids (n=11) were extracted using the AllPrep DNA Mini Kit. For patients 2 and 4, in addition to the organoid lines (PTC-2-I_O and PTC-4-I_O) derived from tumor sites close to the parental tumors (PTC-2_T and PTC-4_T), we also established organoid lines (PTC-2-II_O and PTC-4-II_O) from tumor sites distant from the parental tumors. Accessibility The clustering of the index-coded samples was performed on a cBot Cluster Generation System using the HiSeq PE Cluster Kit (Illumina) according to the manufacturers instructions. In PTC-4, ER and GPER, but not ER, were expressed in tumors and the paired organoids (Supplementary Fig. Herein, we describe our successful creation of 43 tumor organoid cultures and nine adjacent normal tissue organoid cultures derived from patients with endometrial or ovarian cancer. 12634-010] containing 1% HEPES [N-2-hydroxyethylpiperazine-N-2-ethane sulfonic acid; Gibco, No. Disclaimer. All rights reserved. 5C). Xu C, Yuan X, Hou P, Li Z, Wang C, Fang C, Tan Y. Overall, our studies using ER-selective agonists and antagonists demonstrate that ER is the predominant mediator for E2-induced cell proliferation in PTC organoid lines. Threedimensional (3D) organoid culture systems are emerging as potential reliable tools to investigate basic developmental processes of human disease, especially cancer. Objective: Karkampouna S, La Manna F, Benjak A, Kiener M, De Menna M, Zoni E, Grosjean J, Klima I, Garofoli A, Bolis M, Vallerga A, Theurillat JP, De Filippo MR, Genitsch V, Keller D, Booij TH, Stirnimann CU, Eng K, Sboner A, Ng CKY, Piscuoglio S, Gray PC, Spahn M, Rubin MA, Thalmann GN, Kruithof-de Julio M. Nat Commun. Using a 3D organoid system, we report success in long-term culture of prostate cancer from biopsy specimens and circulating tumor cells. Dong Chen and Yawen Tan contributed equally to this work. Recently, Sondorp et al (32) reported the establishment of normal thyroid organoids with a Wnt3a condition medium. The .gov means its official. Similarly, the PTC-6-A/B_O line harboring the BRAF mutation represents the resistance to BRAFV600E-target drugs, highlighting the value of the combination of WES and organoid-based drug-screening assays in PTC organoids. A study comparing drug responses of patient-derived gastrointestinal cancer organoids with the responses of patients in clinical trials is very encouraging (15). Control-FREEC version 11.4 was used to display the distribution of somatic copy number variations (CNVs). Actually, we have tried to generate normal thyroid organoids in Wnt3a condition medium using nontumor thyroid tissues from 5 patients. Two or 3 random pieces were snap-frozen and stored at 80 C for DNA isolation, 2 random pieces were fixed in formalin for histopathological analysis and immunohistochemistry, and the remaining tissues were dissociated and processed for organoid derivation. Each individual specimen was split into several parts that were processed for organoid culture, histology diagnosis, and genomic analysis (Fig. 2019M663101). B, Heat map of gene mutation variations in driver genes of parental tumors and the corresponding organoids. An organoid is a self-organized 3D tissue that is typically derived from stem cells (pluripotent, fetal or adult), and which mimics the key functional, structural and biological complexity of. Patient-derived cancer organoids provide a better reflection of native tumors and may fill in the gap between drug screening based on traditional 2D cancer cell lines and clinical trials. Before The sequencing data were filtered by adaptor and low-quality reads by fastP (version 0.12.6). To determine whether cell proliferation of organoids will respond to estrogen stimulation and to study estrogen receptors that are involved in this process, we treated PTC organoid lines derived from female patients with E2, either in the presence or absence of the selective ER antagonists, or with the selective ER agonists. Notably, increased ER expression has been reported in PTC tissues and seems to correlate with larger tumors and worse clinical features (39). Tumor organoids were subsequently subjected to histological characterization, DNA sequencing, drug screen, and cell proliferation assay, respectively. We observed a significant decrease both in the number and size of organoids formed on omission of EGF or SB202190 (Fig. ab39742) were applied to sections and incubated at 4 C overnight. HY-107216), and G15 (GPER-selective antagonist, No. D Marker expression analysis of papillary thyroid cancer (PTC) organoids. By performing immunofluorescence imaging of a patient-derived, ovarian carcinosarcoma organoid culture, Bi et al. doi: 10.1136/jitc-2022-006290. 4C and 4D, Supplementary Table 3 and 4 [20]). Thyroid cancer has been the most frequent endocrine malignancy with an increasing prevalence in the last decade all over the world (1). Broutier L, Mastrogiovanni G, Verstegen MM, Francies HE, Gavarr LM, Bradshaw CR, Allen GE, Arnes-Benito R, Sidorova O, Gaspersz MP, Georgakopoulos N, Koo BK, Dietmann S, Davies SE, Praseedom RK, Lieshout R, IJzermans JNM, Wigmore SJ, Saeb-Parsy K, Garnett MJ, van der Laan LJ, Huch M. Nat Med. A Establishment of papillary thyroid cancer organoid lines from clinical specimens. Zeng official website and that any information you provide is encrypted On morphological and histopathological analysis, organoids that developed using nontumor tissues demonstrated high similarity to the PTC tumors/organoids and huge differences from the corresponding nontumor thyroid tissues (Supplementary Fig. Groups sharing the same alphabets (a, b) above the bars are not significantly different from one another (P>.05), whereas those not sharing the same alphabets represent significant difference at P less than .05 by ANOVA plus Tukey multiple comparison test. 1A). Surgically resected PTC primary tissues were dissociated and processed for organoid derivation. Matano Dong Chen and others, Organoid Cultures Derived From Patients With Papillary Thyroid Cancer, The Journal of Clinical Endocrinology & Metabolism, Volume 106, Issue 5, May 2021, Pages 14101426, https://doi.org/10.1210/clinem/dgab020. JCYJ20200109120016553), Sanming Project of Medicine in Shenzhen (grant No. 4E). SZSM202011017), and the China Postdoctoral Science Foundation (grant No. 36, 51-54 Yuan et al. Robust organoid models will play a major role in the translation of basic cancer research into novel treatment strategies for individual patients, even in the absence of molecular bases. HHS Vulnerability Disclosure, Help The primary histological types of thyroid cancer include papillary thyroid cancer (PTC), follicular thyroid cancer, Hrthle cell thyroid cancer, medullary thyroid cancer, and anaplastic thyroid cancer. These cultures were generated from numerous cancers, various biopsy sample types, and in different clinical settings. Published by Oxford University Press on behalf of the Endocrine Society. In PTC-6_O, incubation with E2 or PPT could significantly enhance cell proliferation of organoids (Fig. PTC, papillary thyroid cancer; NTG, nodular thyroid goiter; Normal, nontumor normal tissue; T, tumor; O, organoid. Not surprisingly, both point mutation types and contribution of signatures in the parental tumors were well maintained in organoid cultures. TNM staging was performed based on the eighth edition of the American Joint Committee on Cancer staging system. Overall, these results demonstrate that tumor organoids recapitulate the histological features and marker expression of their original tumors. Furthermore, PTC organoids developed in our study showed both cohesive and discohesive structures in the same culture, with the cohesive shapes accounting for the vast majority in all cultures. Deng Zhang Abstract and Figures Osteosarcoma (OS) is the most common primary bone malignancy with high rates of recurrence and metastasis. The whole fluorescence intensity (FI) of EdU and Hoechst 33342 in each well was measured by the VICTOR Nivo Multimode Plate Reader (PerkinElmer Inc), and the proliferation levels of PTC organoids were presented by percentage of EdU-FI/Hoechst-FI. In accordance with previous reports regarding a variety of tumor types (33), PTC organoids also retain a remarkable intratumor mutational heterogeneity of the originating tumors. Genetic characterization of papillary thyroid cancer (PTC) organoids. We then verified whether the somatic mutations of PTC organoid could be maintained in long-term culture. Bookshelf Prostate cancer patient-derived organoids: detailed outcome from a prospective cohort of 81 clinical specimens. ER, ER, and GPER were reported to be expressed in thyroid cancer tissues and/or cell lines (37, 38). These proteins have been considered as potential markers for the clinical diagnosis and laboratory test of PTC. , Bonofiglio D, Albanito L, et al. , Bayram T, van de Wetering M, et al. Historically, the most universal preclinical models were mouse models and patient-derived cell lines propagated in 2-dimensional (2D) culture. HY-100689), DPN (2,3-bis(4-hydroxyphenyl)-propionitrile; ER-selective agonist, No. Epub 2014 Sep 23. Loss of tumor suppressors PTEN, TP53, RB1 , and CDKN2A in prostate organoid, Figure 6. A, Flow diagram of establishment and characterization of PTC organoids, including the generation of PTC organoid lines from patients undergoing surgery using optimized medium components, as well as the histological characterization, DNA-sequencing, drug screen, and cell proliferation assay on PTC organoids. Breast cancer is a complex disease that has been classified into several different histological and molecular subtypes. A, Heat map of logIC50 values for 24 anticancer drugs used to treat 11 PTC organoids derived from 8 patients by applying nonlinear regression. Indeed, the past decade has seen an evident trend toward the use of organoids derived from healthy and malignant tissues. Pictures of organoids were taken using an Olympus IX73 microscope. A. Quantification of number of non-synonymous single nucleotide variations (SNV, pink) and insertion-deletions (Indel, orange) in each organoid sample compared to average total SNV and Indels of published whole-exome datasets (green). The PDOs can be used to build a biobank and perform single-cell sequencing, drug screening and genetic analyses. Patient-Derived Cancer Organoid Cultures to Predict Sensitivity to Chemotherapy and Radiation. T Organoids derived from MPT may be a novel preclinical model for testing personalized therapies for patients with MPT. In contrast, DPN and G-1 had no effect. Organoid Culture. Drug responses in papillary thyroid cancer (PTC) organoid lines. A, Heat map of gene mutation variations in the most significantly mutated genes of PTC tumors and the matched organoids. 17504-044), N-acetyl-L-cysteine (1.25 mM, Sigma-Aldrich, No. Here, we discuss the applications of patient-derived tumor organoid cultures and the possibilities in using complex immune-organoid cultures to provide preclinical testing platforms for precision cancer immunotherapy. Mutational landscape of patient-derived prostate cancer organoid lines. However, this is starting to change with the development of 3D cultures of developing tissues, called organoids. For passaging, organoids were resuspended in 5-mL TrypLE Express with incubation at 37 C for approximately 5 minutes. Stem Cell Reports. Left behind was a cancer organoid - a 3D mass . When required, the organoids were thawed using standard thawing procedures and cultured as described earlier. Since 2018, 17 publications have examined PDOs as a potential predictive biomarker in the treatment of cancer. M See also Supplementary Table 4 (20). An official website of the United States government. Disclosures: The authors have nothing to disclose. 2022 Nov 21;10:1043117. doi: 10.3389/fcell.2022.1043117. 2019 Dec;52(4):181-186. doi: 10.1007/s00795-019-00217-6. Preclinical Models of Follicular Cell-Derived Thyroid Cancer: An Overview from Cancer Cell Lines to Mouse Models. To understand thyroid cancer and translate this knowledge to clinical applications, patient-derived PTC organoids as a promising new preclinical model were established. Cancer cell lines and patient-derived xenograft (PDX) models are widely used as preclinical tumor models for screening drugs and assessing drug responses. Zhao Z, Chen X, Dowbaj AM, Sljukic A, Bratlie K, Lin L, Fong ELS, Balachander GM, Chen Z, Soragni A, Huch M, Zeng YA, Wang Q, Yu H. Nat Rev Methods Primers. PTC, papillary thyroid cancer; NTG, nodular thyroid goiter. Published by Oxford University Press on behalf of the Endocrine Society. sharing sensitive information, make sure youre on a federal Organoids were passaged at a 1:2 to 1:4 dilution every 2 weeks. Similar morphological features were also found in the organoid lines of breast cancer (12) and pancreatic cancer (31). A co-clinical trial for the treatment of locally advanced rectal cancer has demonstrated that tumor organoids can predict patient chemoradiation responses in the clinic (16). Before To prepare frozen stocks, organoid cultures were dissociated and mixed with Recovery Cell Culture Freezing Medium (Gibco, No. Mutation effect predictions and annotations were performed by ANNOVAR and dbSNP. 6C). All data generated or analyzed during this study are included in this published article or in the data repositories listed in References (20). Estradiol was shown to promote cell proliferation of PTC organoids in the presence of estrogen receptor (ER), regardless of the expression of ER and G proteincoupled ER. 6A, Supplementary Fig. Results from all these studies suggest that ER is the predominant mediator for the E2-induced cell proliferation in organoid lines. These data suggest that mutations in the RAS/BRAF/MAPK pathway alone were insufficient to compensate for the absence of EGF, and additional molecular variations in EGF signal pathways may be required. Copyright 2014 Elsevier Inc. All rights reserved. Loss of tumor suppressors PTEN,, Figure 5. 2 [20]). For instance, we observed that PTC-1_O and PTC-2_O appeared as dense spheres consisting of multiple layers of cells, whereas PTC-3_O exhibited a hollow morphology with cells that appear to be wreathed (Fig. The expression of ER, ER, and GPER in PTCs from women (patients 2, 3, 4, 6, and 8) and their derived organoids was first examined by immunohistochemical staining (Fig. 7074S) or horseradish peroxidaselinked antimouse immunoglobulin G (1:1000; Cell Signaling Technology, No. Use of conditional reprogramming cell, patient derived xenograft and organoid for drug screening for individualized prostate cancer therapy: Current and future perspectives (Review). PPT, DPN, and G-1 are widely used agonists for ER, ER and GPER, respectively. A and E, Immunohistochemical staining of ER, ER, and G proteincoupled estrogen receptor (GPER) in tumor-organoid pairs of PTC-6 and PTC-8, respectively. 5). SPOP and FOXA1 mutations are associated with PSA recurrence in ERG wt tumors, and SPOP downregulation with ERG-rearranged prostate cancer. The advent of cancer organoids has allowed researchers to overcome limitations of the existing models by means of their stable culture characteristics, flexible manipulation, and faithful recapitulation of the patient-specific signature and genetic landscape of the primary tumors, especially tumor heterogeneity (26). After preincubation for 24 hours, a 6-point, 5-fold dilution series of each compound as well as DMSO controls were added. 2017 Dec;23(12):1424-1435. doi: 10.1038/nm.4438. The cell pellets were resuspended in ice-cold Matrigel (Corning, No. After 2 days of treatments, cell proliferation of PTC organoids was measured by 5-ethynyl-2deoxyuridine (EdU) uptake according to the manufacturers instructions. The emergence of organoid technology has provided a new direction for colorectal cancer research. C, Representative scatterplots of 1 area under the curve (AUC) values from 2 biological replicates (different passages) of the drug screening data. From 2009 to 2021, use of organoid technology has been rapidly increasing in cancer research, especially for therapeutic screening and precision medicine (18-20). G9683) according to the manufacturers instructions, and luminescence was detected in the VICTOR Nivo Multimode Plate Reader (PerkinElmer Inc). Trypsinization was stopped by the addition of 5 mL AdDMEM/F12+++ medium containing 20% fetal bovine serum (FBS; Gibco, No. The success rate for the generation of organoid lines was 71.4% for PTC (10 of 14 PTC tissues) and 75% for NTG (3 of 4 NTG tissues). Roerink Patient-derived xenografts and organoids model therapy response in prostate cancer. These organoid lines have been propagated by serial passaging and have been successfully cryopreserved and retried. The corresponding patients normal blood was also sequenced and used as a reference. E, Representative bright-field images of PTC- and NTG-derived organoids from 3 patients. To assess whether PTC organoids faithfully recapitulate the genomic profiles of their originating tumors, we next subjected DNA from 11 PTC organoid lines and 8 parental tumor tissues to WES. Results: We demonstrate that patient-derived cancer organoids represent the cancers from which they were derived, including key histologic and molecular features. 3603), in duplicates. The whole fluorescence intensity (FI) of EdU and Hoechst 33342 in each well were measured using the VICTOR Nivo Multimode Plate Reader (PerkinElmer Inc), and the proliferation levels were presented as a percentage of EdU-FI/Hoechst-FI. 2). Whole-exome sequencing shows a low mutational burden, consistent with genomics studies, but with mutations in FOXA1 and PIK3R1, as well as in DNA repair and chromatin modifier pathways that have been reported in advanced disease. ab75635), ER (1:300, Abcam, No. See also Supplementary Table 2 (20). Passage numbers of the organoid lines were PTC-1_O, P4; PTC-2-I_O, P5; PTC-2-II_O, P5; PTC-3_O, P6; PTC-4-I_O, P4; PTC-4-II_O, P4; PTC-5_O, P6; PTC-6-A_O, P2 (culturing for 26 days); PTC-6-B_O, P8 (culturing for 121 days); PTC-7_O, P6; PTC-8_O, P6. 5 and Supplementary Fig. Emerging organoid-immune co-culture models for cancer research: from oncoimmunology to personalized immunotherapies. Showing numerous advantages over 2D cell lines, 3D cell lines, and primary cell culture, organoids have been applied in drug screening to demonstrate the correlation . These results were consistent with the previous finding that CRISPR (clustered regularly interspaced short palindromic repeats)engineered KRASG12V organoids gained independence of EGF but required EGF for their optimal growth (28). Time-lapse phase-contrast imaging of the cohesive PTC organoids showed that many organoids seem to be clonally derived (Fig. Because all the patients who had undergone surgery between July 2019 and June 2020 were alive without disease progression at the time of our manuscript preparation, we cannot compare the organoid response to drugs with patients clinical outcomes. Keywords: PMC Tumor organoids were subsequently subjected to histological characterization, DNA sequencing, drug screen, and cell proliferation assay, respectively. These primary cell cultures, called organoids, are three dimensional stem-cell derived cultures that support the propagation of phenotypic, genetic and transcriptomic characteristics from the original tissue and retain the self-renewal properties of stem cells and their ability to undergo multilineage differentiation indefinitely. Federal government websites often end in .gov or .mil. 100-26), R-spondin-1 (500 ng/mL, Peprotech, No. G All rights reserved. sharing sensitive information, make sure youre on a federal Drug sensitivity testing and organoid cell . Summary of organoid establishment, sources and applications. , Ogundipe VML, Groen AH, et al. Drug sensitivity assays of PTC organoids demonstrated patient-specific drug responses (Fig. The growth and passage of PTC and NTG organoid lines were recorded (Fig. Passage numbers of the organoid lines were PTC-6_O, P4; PTC-8_O, P7. D, Effects of 100 nM E2, in the presence or absence of 1 M fulvestrant, tamoxifen citrate, MPP (1,3-bis(4-hydroxyphenyl)-4-methyl-5-[4-(2-piperidinylethoxy)phenol]-1H-pyrazole), PHTPP (4-(2-phenyl-5,7-bis(trifluoromethyl)pyrazolo[1,5-a]pyrimidin-3-yl)phenol), or GPER-selective agonist G15, on cell proliferation of PTC organoid lines. Q Epub 2019 May 15. Huang Online ahead of print. See also Supplementary Figure 1 (20). In addition to histological conservation, subsequent marker expression analysis revealed that organoids and their matched tumors exhibited similar staining patterns for CK19, galectin-3, Tg, Ki-67, CD117, and TTF-1 (see Fig. Papillary thyroid cancer (PTC) has been one of the most frequent endocrine malignancies around the world. After cluster generation, the DNA libraries were sequenced on an Illumina HiSeq platform and 150bp paired-end reads were generated. Abbreviations: F, female; M, male; NA, not available; NTG, nodular thyroid goiter; PTC, papillary thyroid cancer. Here, we demonstrate that the most common genetic mutations and mutational signatures in human PTC tissues were also observed in organoid cultures. HY-103456), G-1 (G proteincoupled estrogen receptor [GPER]-selective agonist, No. 8600 Rockville Pike All experiments were repeated 3 times in duplicates, and results were shown as means from three independent experiments. Nat Methods. N0636), noggin (100 ng/mL, PeproTech, No. S7067). Drug concentrations ranged from 3.2104 M to 1 M, from 3.2103 M to 10 M, or from 3.2102 M to 100 M, depending on the individual drug properties. It furthers the University's objective of excellence in research, scholarship, and education by publishing worldwide, This PDF is available to Subscribers Only. , Chen GG, Vlantis AC, van Hasselt CA. We observed both cohesive (dense and solid structures) and discohesive (irregularly shaped structures) organoids within the same culture derived from the same specimen (Fig. Abbreviations: Akt, protein kinase B; BRAF, B-Raf proto-oncogene, serine/threonine kinase; Cat., catalog; MEK, mitogen-activated protein kinase kinase; mTOR, mammalian target of rapamycin; PDGFR, platelet-derived growth factor receptor ; VEGFR, vascular endothelial growth factor receptor. Detailed information for each drug can be found in Table 2. A9165), nicotinamide (10 mM, Sigma-Aldrich, No. Prostate cancer organoids exhibit diverse, Figure 4. . , Mastrogiovanni G, Verstegen MM, et al. Although most PTC patients have a favorable prognosis, a subgroup of patients die, especially when disease recurrence occurs. In principle, mutations in the RAS/MAPK pathway, including KRAS and BRAF, were expected to be EGF independent. Epub 2022 Dec 26. Relationship between genetic alterations and clinicopathological characteristics of papillary thyroid carcinoma. 4E and Supplementary Table 5 [20]). Generally established from both healthy and diseased tissue, adult stem cell-derived organoids consist of epithelial cells and provide an opportunity to expand patient-derived cells in the. 3 [20]). The majority of biomedical research efforts on PTC has so far centered on using a limited number of thyroid cancer cell lines. Scale bar, 100 m. The images shown are from PTC-1_O as a representative sample. Dissociated tissues were spun down at 300g for 5 minutes, washed once with AdDMEM/F12+++ medium (Advanced DMEM [Dulbeccos modified Eagles medium]/F12 medium [Gibco, No. Unfortunately, normal thyroid organoid cultures have not been established from nontumor thyroid tissues in this study. Drug sensitivity assays of PTC organoids demonstrate patient-specific drug responses, and large correlations with the respective mutational profiles. Cancer patient-derived organoids from Japan's Fukushima Medical University have become the latest technology to be made available via CancerTools.org. SOP40104: Cryopreservation of Patient-Derived Organoid (PDOrg) Cultures; SOP30108: Implantation of Patient-Derived in vitro Material (2D and 3D cell cultures) for Generation of Cell Line Xenografts (CLXs) PDMR Genomics. This site needs JavaScript to work properly. government site. A synopsis of prostate organoid methodologies, applications, and limitations. From this experiment, we found that the addition of EGF and SB202190 are sufficient for the formation and growth of human PTC organoids (Supplementary Fig. Neither PHTPP nor G15 inhibited basal and E2-induced cell proliferation (see Fig. Similar results were also observed in PTC-2-I, PTC-3, and PTC-4-I organoid lines (see Supplementary Fig. 6B and 6C). Tissues were resuspended in 5 mL of TrypLE Express (Gibco, No. J Mol Med (Berl). Cancer cell lines do not generally capture the 3D organ structure and original heterogeneity, so they are fundamentally limited in representing the complexity of cancer (4).
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