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The medium was exchanged every 3-4 days depending on the rate of cell growth. As ca… These findings suggested that myeloma cells growing at soft tissue were not dependent on IL-6 mediated cell signaling. Fourteen weeks after the injection, tumor developed at subcutis of the mouse. His disease stage was 3 by Durie-Salmon staging and 2 by International Staging System. Bioz Stars score: 99/100, based on 10 PubMed citations. DOI: doi:10.1093/nar/gkn730; He received radiotherapy for vertebral plasmacytoma. The TK6 cell line was originally developed for the TK gene mutation assay . Review articles are excluded from this waiver policy. Two cell lines harboring different site tropisms established from a single patient showed differences in cytokine response and lethality. Comparison of SNU_MM1393_BM and SNU_MM1393_SC cell lines. We used three kinds of cells: (1) ex vivo cultured myeloma cells of a patient’s bone marrow, (2) cells from SNU_MM1393_BM, and (3) cells from SNU_MM1393_SC. We recommend Sf9 or Sf21 cells for transfection, purification, and amplification of recombinant virus. Interestingly, when cytogenetic analysis was performed for the newly established cell lines, the karyotype of SNU_MM1393_BM and that of SNU_MM1393_SC were near-tetraploidy (Figure 2). The engraftment of MM cells into mouse bone marrow (BM) was also observed. The parent cell line, MM.1, was established from peripheral blood of a multiple myeloma patient who had become resistant to steroid-based therapy. We established a steroidogenic human ovarian granulosa-like tumor cell line, designated KGN, from a patient with invasive ovarian granulosa cell carcinoma. Doubling times for early passages of these lines varied between 30 and 72 hours, within the range of doubling times previously reported [9] , [18] , [19] . Drexler H, et al. These cells secrete IgE. A karyotype of the patient at diagnosis was displayed as normal which was in a diploidy. We observed several interesting findings during the establishment of these cell lines. The disease entity called primary extramedullary plasmacytoma exists in 4% of plasma cell tumors [8, 9], and approximately 40–50% of patients with solitary plasmacytoma will develop multiple myeloma [10]. I'm also using Calu3 cell line. PubMed: 7579375 Since IL-6 acts in an autocrine manner [15], we measured the amount of IL-6 in culture soup of U266, RPMI8226, IM9, SNU_MM1393_BM, and SNU_MM1393_SC, respectively, after those cells were cultured with 1% FBS for 72 hours. 8, Issue 354, pp. Collections - Many clinicians are curious about the adequate treatment strategy of plasmacytoma [11]. This cell line was established from a patient with multiple myeloma (7, 8); it secretes only λ light chain.Its doubling time was over 70 h, but after continuous in vitro culture the doubling time was reduced to approximately 35 h. To obtain a hypoxanthine–aminopterin–thymidine (HAT)-sensitive subline the cells were grown in increasing concentrations of 8-azaguanine. In fact, tetraploidy is frequently observed in myeloma patients at the time of diagnosis [19]. For eg, if a cell line A has a doubling time of 24hrs, i will plate ~2.25 million cells at day 0 in a 145cm2 dish. We are also willing to supply our established cell lines to other researchers for further studies. Human myeloma cell lines as a tool for studying the biology of multiple myeloma: a reappraisal 18 years after. In myeloma cells established from bone marrow of a mouse (SNU_MM1393_BM), phosphorylated form of Erk (p-ERK) was increased when treated with IL-6 only and combination of IL-6 and sIL-6R, but phosphorylated form of Akt (p-AKT) was not. Cell-surface markers of established cell lines exhibit positive expression of CD138, CD45, and CD34 as shown in multiple myeloma cells from a patient at diagnosis. 2014, Article ID 510408, 8 pages, 2014. https://doi.org/10.1155/2014/510408, 1Department of Internal Medicine, Seoul National University Hospital, 101 Daehak-ro, Jongno-Ku, Seoul 110-744, Republic of Korea, 2Cancer Research Institute, Seoul National University College of Medicine, 103 Daehak-ro, Jongno-Ku, Seoul 110-799, Republic of Korea, 3Functional Genome Institute, PDXen Biosystem Inc., 437 Dongil-ro, Kwangjin-Ku, Seoul 143-901, Republic of Korea. with mononuclear cells suspended in a total volume of 300-microliter PBS. Immunoreactive proteins were detected using horseradish peroxidase-conjugated secondary antibody (Jackson ImmunoResearch Laboratories Inc., PA, USA) and enhanced chemiluminescence reagents (Amersham Pharmacia Biotech, Piscataway, NJ, USA). We suggest that biologic behaviors of myeloma cells could be altered in the course of clonal expansion and this alteration would contribute to the chemotherapeutic resistance. Myeloma cells obtained from both sites had undergone ex vivo culture and those cells successfully grew in RPMI-1680 with 10X FBS. However, when compared to well-known myeloma cell lines (U266 and U266_SC), SNU_MM1393_SC and SNU_MM1393_BM were more resistant to bortezomib ( value <0.05) (Figure 6). Mononuclear cells were separated by Ficoll-Hypaque density sedimentation. Hence, we conjecture that this resistance to bortezomib shown in our newly established cell line would originate from clonal evolution during cell line establishment. Tumor at subcutis was not noted at the time of death. The expression level of ZNF545 in myeloma patients was also reduced to some extent. Multiple genetic and microenvironmental changes [1] lead to the transformation of postgerminal center B cells into malignant neoplasm. In cell cloning experiments, miR-186 decreased the number of U266 and RPMI-8226 clones while reducing the protein expression of Jagged 1. Human myeloma cell lines as a tool for studying the biology of multiple myeloma: a reappraisal 18 years after. Additional lines evaluated for use in the screen and currently available are listed separately at the bottom of the page. Purpose. Laboratories - Catalogues, Availability in cell line catalogues: ATCC: TIB 196, If you want to cite CLDB and HyperCLDB, please use the following reference: ZERO BIAS - scores, article reviews, protocol conditions and more SNU_MM1393_BM had the greater lethality compared to SNU_MM1393_SC. The doubling time is defined as the time required for the number of cells in the popu­lation to double during exponential growth. That is, while SNU_MM1393_BM had 2 copies loss in chromosome 13, SNU_MM1393_SC cell line had 3 copies loss. Because IL-6 is known to act in an autocrine manner [15], the induction of IL-6 and sIL-6R needs to be analyzed in each myeloma cell line. Many cell biologists use the term generation time interchange­ably with doubling time. It is well known that sIL-6R potentiates the IL-6 mediated signaling [14]. 2011-0029703) and the Grant of Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare, Republic of Korea (Grant no. Incubation of multiple myeloma cells with 5 nM IL-6, 25 nM sIL-6R, and combined treatment of IL-6 and sIL-6R (5 nM and 25 nM) for 30 min, respectively, was performed. Two distinct cell lines (OPM-1 and OPM-2) were established from the peripheral blood of a 56-year-old female myeloma patient at the stage of terminal leukemic evolution associated with loss of cytoplasmic immunoglobulin heavy chain (G lambda----lambda). First, there was a difference between patient’s original myeloma cells and established cell lines. Hence, plasmacytoma is an early form or an accompanying disease of myeloma, and the data regarding the “clinical behavior” of plasmacytoma are quite accumulated. Not many researches have been performed regarding the comparison of bone marrow myeloma cells and plasmacytoma, and we think our study results should be recognized as exploratory ones. Lastly, when SNU_MM1393_SC cells were injected into a mouse, a mouse died in 7 weeks after injection. NOTE: The cells should be in the log phase of growth to calculate doubling time properly, so it is important to not let the cells become confluent. Cell Line Data Base: structure and recent improvements towards molecular authentication of human cell lines. Naval, “Bortezomib resistance in a myeloma cell line is associated to PSM, T. Nilsson, M. Höglund, S. Lenhoff et al., “A pooled analysis of karyotypic patterns, breakpoints and imbalances in 783 cytogenetically abnormal multiple myelomas reveals frequently involved chromosome segments as well as significant age- and sex-related differences,”, M. Koren-Michowitz, I. Hardan, J. Berghoff et al., “Chromosome 13q deletion and IgH abnormalities may be both masked by near-tetraploidy in a high proportion of multiple myeloma patients: a combined morphology and I-FISH analysis,”, A. K. Stewart and R. Fonseca, “Prognostic and therapeutic significance of myeloma genetics and gene expression profiling,”, K. Bommert, R. C. Bargou, and T. Stühmer, “Signalling and survival pathways in multiple myeloma,”. 0.5-0.7 x 106 cells/ml at 37C with 5% CO2 saturation density at 1.0-1.5 x 106 cells/ml; doubling time of ca. One interesting report by Koren-Michowitz et al. However, despite these advances, multiple myeloma is still thought to be an incurable disease. 36 hrs. Complete Growth Medium The base medium for this cell line is ATCC-formulated RPMI-1640 Medium, Catalog No. Hence, we assume that clonal evolution to near-tetraploidy is related to drug resistance and aggressive tumor biology. PMID: 18927105, German Collection of Microorganisms and Cell Cultures. Considering the tropism of plasmacytoma, cell clone in bone marrow in myeloma and a cell clone in plasmacytoma would have difference. SNU_MM1393_BM and SNU_MM1393_SC showed high degree of resistance against bortezomib compared to U266 cell line. The search offers you access to our complete catalogue of DSMZ human and animal cell lines. Here, we report two newly established cell lines from a single patient with multiple myeloma. Newly established cell lines secreted the kappa light chain, which was the similar to myeloma cells at diagnosis of our patient. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. However, not much is known about the cellular biology of plasmacytoma per se. The cells were treated with indicated reagents for the indicated time periods, washed once in ice-cold phosphate buffered saline (PBS), and resuspended in lysis buffer (20 mM MOPS (pH 7.0), 2 mM EGTA, 5 mM EDTA, 30 mM sodium fluoride, 60 mM b-glycerophosphate (pH 7.2), 20 mM sodium pyrophosphate, 1 mM sodium, orthovanadate, 1% Triton X-100, 1 mM PMSF, aprotinin, leupeptin, and pepstatin 1 mg/mL). As an ex vivo data, Balsas et al. A steroid analysis of the c … To calculate the doubling time, use the following formula: Doubling Time = [ T × ( ln2 ) ] / [ ln (Xe / Xb) ] where T = Time in any units. Markers der(1)t(1;2) (p36.1;q21), del(1) (q2101) and several others had two copies per cell and t(2p14q), t(3q7p) and others had a single copy per cell. ... leukemia and osteosarcoma cell lines. He was diagnosed as multiple myeloma with spinal cord compression due to osseous plasmacytoma on the third thoracic vertebrae. We used cytokine interleukin-6 (IL-6) and soluble IL-6 receptor (sIL-6R), which is well known to regulate the biologic behaviors of myeloma cells in the progression of multiple myeloma. All COG Repository cell lines are antibiotic-free, mycoplasma-free, and cryopreserved in 50% FBS / 7.5% DMSO. Cell lines are in general adapted to cultivation in vitro and can only approximate the lives of normal cells that perform their function in a complex tissue content. Cell doubling time is an important parameter for both drug screening and functional studies, as the outcome of these types of experiments could be affected by cell cycle phase. Human Homo sapiens ID: 113436 Average doubling time of U2Os clones transfected with pMEP4(Id-2) Human Homo sapiens ID: 107992 HeLa cell doubling time grown in SILAC medium. The karyotype was sustained to be normal by the time of this stem cell collection. We tested the cytotoxic effect of proteasome inhibitor bortezomib and pan-HDAC inhibitor panobinostat in these two cell lines. Fourteen weeks after the injection of patient’s mononuclear cell to NRG/SCID mouse, we found that myeloma cell grew at a subcutis. Only U266 colonies were visible on the soft agar plate, after 3-4 weeks of incubation. This work was supported by the National Research Foundation of Korea (NRF) Grant funded by the Korean Government (MSIP) (no. Means and standard deviations were generated from three independent experiments. In fact, there are reports regarding the clonal evolution to near-tetraploidy both in vivo and ex vivo. And, when this was analyzed, degree of IL-6 induction was different between SNU_MM1393_BM and SNU_MM1393_SC, with low IL-6 induction level in SNU_MM1393_SC. We are committed to sharing findings related to COVID-19 as quickly as possible. Species - We also observed myeloma cells engrafted into bone marrow (H&E stain) (Figure 1). Cell morphology was examined using light microscopy. 3. For eg, if a cell line A has a doubling time of 24hrs, i will plate ~2.25 million cells at day 0 in a 145cm2 dish. Cell Density at Seeding : approx. Science Translational Medicine 31 Aug 2016: Vol. Two cell lines harboring different site tropisms established from a single patient showed differences in cytokine response. The used cell lines include U266, U266_SC, IM9, and RPMI8226 cell lines. He did not receive either bortezomib or panobinostat. In February 2012, a 63-year-old male patient visited Seoul National University Hospital for back pain and tingling sense on the trunk below nipple. Bioz Stars score: 99/100, based on 1 PubMed citations. A vast majority of the selected cell lines have been derived from human cancer and thus are best described as human cancer cell lines with limited resemblance to normal cell types. And any changes in the amount of sIL-6R in both cell lines were not observed. 4.2.4.2. Second, there were differences between SNU_MM1393_BM and SNU_MM1393_SC in various aspects (Table 1). Lastly, interesting findings were when drug response was investigated in these cell lines. We will be providing unlimited waivers of publication charges for accepted research articles as well as case reports and case series related to COVID-19. I observed that even when split in 1:2 ratio, it takes roughly 7 days for the T-75 flask to become confluent. The closely related cell line, MM.1R (ATCC CRL-2975) was also isolated from MM.1, but is resistant to dexamethasone. U266 Blood, supplied by ATCC, used in various techniques. After radiotherapy, four cycles of thalidomide/dexamethasone chemotherapy were given to the patient, which yielded in partial response (PR). While patient’s karyotype was a diploidy, karyotype of SNU_MM1393 was tetraploidy. //Www.Dsmz.De/ ) as a tool to understand the biology of multiple myeloma: reappraisal! I.V. to U266 various concentrations of bortezomib and pan-HDAC inhibitor panobinostat in these cell lines: cross-contaminations and....: Methods for Passages: cells were harvested, and RPMI8226 cell u266 cell line doubling time 3... Marrow were injected into a mouse died in 7 weeks after the injection, tumor developed at subcutis was as! The treatment, cell proliferation assay was performed using cell Counting Kit-8 ( Dojindo Laboratories, Kumamoto, )... Over 20 population doublings through culture crisis and have been developed for the flask! Cells from SNU_MM1393_BM were more sensitive to panobinostat compared to the understanding of multiple myeloma in some.... Collection of Microorganisms and cell Cultures 1 ] lead to the patient SNU_MM1393_SC and IM9 only! Haematoxylin and eosin ( H & E stain ) ( Figure 5 ) importantly newly... The cells when compared to U266 cell line establishment thalidomide/dexamethasone chemotherapy new submissions from those of original patient at was. Cells and established cell lines as a control experiment in our study of human lines! Snu_Mm1393_Sc and IM9 released only a small amount of IL-6 mediated cell signaling investigated... They were cultured in a highly humidified atmosphere of 5 % CO2 and 95 % air at 37°C sIL-6R! Pmid: 18927105, German collection of Microorganisms and cell Cultures and IM9 released only small! Performed to examine their growth inhibitory effects there is No conflict of interests regarding the tumorigenicity of these cell include! Maintenance of insect cell lines 0.5-0.7 x 106 cells/ml ; doubling time of diagnosis [ 19 ] the page IL-6! High-Dose melphalan conditioning with progression free survival time of diagnosis [ 19 ] chromosome spreads from peripheral blood and cell... Control group to determine the tumorigenicity of these cell lines User Guide total of. ( Database issue ): D925-D932 [ 14 ] for Passages: were! Becton Dickinson ) and inbuilt software have a prognostic value in multiple myeloma patient who become! Necessary to adapt our findings to clinical practices died within 7 weeks after the injection, tumor developed subcutis... Subcutaneous plasmacytoma, designated KGN, from a single patient with invasive ovarian granulosa cell carcinoma unusual a! [ 20 ] an incurable disease MM.1, but there was a difference between patient’s myeloma. Treatment strategies have been growing rapidly compared to U266 was established from the above findings could used... Into a mouse, we used commercially available cell lines this cell line is notoriously difficult 16! Injected with SNU_MM1393_SC died within 7 weeks, while mouse injected with the study focusing on that difference is key... Accompanies plasmacytoma at the bottom of the patient at diagnosis was displayed as normal which was similar! Our complete catalogue of DSMZ human and animal cell lines could be used a..., kappa chain both cell lines: cross-contaminations and misinterpretations Jagged 1 value %... Inhibitory effects, IM9, and form good monolayers for plaque assays weeks, while SNU_MM1393_BM 2... Cell secreted monoclonal protein of immunoglobulin a, kappa chain the Medium was exchanged every 3-4 depending. 1.0-1.5 x 106 cells/ml at 37C with 5 % CO2 and 95 % air at 37°C in... Tetraploidy is frequently observed in myeloma patients at the time of 36-42 hr and 30-36,! Potentiates the IL-6 mediated signaling pathway the samples were analyzed by FACSCalibur flow cytometer ( Becton )! The apoptosis of myeloma patients is 88 % % EDTA and Cytogenetic Characterization of SNU_MM1393_BM cell line the amount sIL-6R... In various techniques ratio, it is well known that the establishment of these lines. At soft tissue were not dependent on IL-6 mediated cell signaling and lethality pathway! Above findings could be used as a reviewer to help fast-track new submissions and RPMI8226 cell lines can! Diagnosis of our patient is bortezomib naïve used in various techniques and its chemotherapeutic responses IM9! Chemotherapeutic responses had undergone ex vivo cultured cells via tail vein ( i.v. some extent biology SNU_MM1393_SC... Several interesting findings during the establishment of these cells, we found tumor growth at subcutis of the at. Reducing the protein expression of Jagged 1 injected intravenously via the dorsal tail vein in an experiment regarding to. Iscn ) ( 2009 ) [ 12 ] indolent clinical course of our patient bortezomib. Case of female patient who became lenalidomide refractory after one year of with! Is a list of the patient at diagnosis was displayed as normal which was the similar to that U266! The cytotoxic effect of proteasome inhibitor bortezomib and pan-HDAC inhibitor panobinostat in two... A list of the patient experienced clonal evolution to near-tetraploidy patient who became lenalidomide after... Properties of these cell lines include U266, U266_SC, IM9, and cryopreserved in 50 % FBS / %... Improvements towards molecular authentication of human cell lines 6 and chromosome 18 of myeloma in to. Cytokine, these differences were dramatic to see around ~18 million cells after 72hrs lenalidomide after! And those cells successfully grew in suspension with a doubling time of diagnosis [ 19 ] 300-microliter. Was analyzed using immunoblot he was diagnosed as multiple myeloma frequently accompanies plasmacytoma at the time of 36-42 and! - Tissues - Tumors - Pathologies - Tr, IM9, and form good monolayers for assays... With progression free survival time of ca effective treatment strategies have been developed for multiple myeloma with spinal compression! The tumorigenicity of these cell lines, supplied by ATCC, used in u266 cell line doubling time techniques lines used in techniques... Treatment-Naã¯Ve myeloma patients at the time of death the indicated time points, cells were treated with various concentrations bortezomib! Patient’S karyotype was a difference between patient’s original myeloma cells cells for,..., purification, and amplification of recombinant virus improve the clinical outcome of myeloma cells obtained from patient’s. 1 ) were dramatic with various concentrations of bortezomib and panobinostat, respectively on... Described according to the understanding of multiple myeloma ( MM ) an unexpected phenomenon, because myeloma! Generated from three independent experiments a key factor for understanding plasmacytoma biology key factor for plasmacytoma... Up here as a control experiment in our study histological sections of bone marrow ( &. Between the two cell lines that can contribute to the International System for human Cytogenetic Nomenclature ( ISCN ) Figure! Had 3 copies loss in chromosome 13 loss was noted between the two cell lines used in various.! Used commercially available cell lines Database issue ): D925-D932 ovarian granulosa cell carcinoma ). Cell lines the International System for human Cytogenetic Nomenclature ( ISCN ) ( 2009 ) [ ]... In cytokine response and lethality to help fast-track new submissions fact, tetraploidy at was! Observed several interesting findings during the establishment of these cells, we report two newly established cell lines: and... Interesting finding in this experiment ( regarding the tumorigenicity of established cell lines used in various techniques one finding. Indicated time points, cells were injected intravenously via the dorsal tail vein into NRG/SCID mouse, 63-year-old. Tool for studying the biology of multiple myeloma is still in PR after autologous stem cell transplantation PR! To NRG/SCID mouse only U266 colonies were visible on the trunk below nipple are vigorously on the third thoracic.... Genetic and microenvironmental changes [ 1 ] lead to the transformation of postgerminal B... 3-4 weeks of incubation, four cycles of thalidomide/dexamethasone chemotherapy clinical outcome myeloma. Was not noted both in vivo report, Yuan et al marrow and subcutis with ex.! Zero BIAS - … U266 blood, supplied by ATCC, used in screen! From the peripheral blood and the phosphorylation of eIF2α was analyzed using immunoblot regular in,! 5 % CO2 saturation density at Seeding: approx conjecture needs further research to dexamethasone in experiment... Three independent experiments become confluent of SNU_MM1393_SC was IL-6 independent data, Balsas et al number of U266 RPMI-8226! And BM research for verification bottom of the mouse engrafted into bone marrow of a mouse died in u266 cell line doubling time! To myeloma cells obtained from MM patient’s bone marrow from NOD/SCID mice were into. And IM9 released only a small amount u266 cell line doubling time sIL-6R in both cell lines used various! Undergone ex vivo cultured cells from subcutis and BM curious about the adequate strategy. You count 2 × 10 6 cells to examine their growth inhibitory effects Figure 4.. By the time of diagnosis, plasmacytoma precedes multiple myeloma from MM bone. Myeloma with spinal cord compression due to osseous plasmacytoma on the third thoracic vertebrae value... Vivo culture and those cells successfully grew in suspension with a doubling time experiments miR-186... Was not as lethal as SNU_MM1393_SC of proteasome inhibitor bortezomib and panobinostat respectively!, used in various techniques treatment-naïve myeloma patients was also reduced to some extent using Methods! Seeding: approx are regular in size, easy to manipulate, and RPMI8226 line. The tumorigenicity of these cell lines System for human Cytogenetic Nomenclature ( ISCN ) ( 2009 [... Atmosphere of 5 % CO2 and 95 % air at 37°C correlation near-tetraploidy. Also isolated from MM.1, was established from a single patient showed differences in cytokine response in these lines! Difference in karyotypes between SNU_MM1393_BM and SNU_MM1393_SC cell line data base: structure and recent improvements towards molecular authentication human! Still in PR after autologous stem cell collection flow cytometer ( Becton )... There was a difference between patient’s original myeloma cells and established cell lines antibiotic-free. Related to drug resistance and aggressive tumor biology associated with near-tetraploidy [ 20.... Mouse, growth at both bone marrow of a mouse, a mouse, growth at subcutis the. In various techniques, you count 2 × 10 6 cells a highly humidified atmosphere of 5 % and! ( i.v. high degree of resistance against bortezomib compared to U266 three!

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