Briefly, K562/S cells were grown initially in the continuous presence of 0.5 × 10 −6 M VBL (K562/R0.5) until the population doubling time was similar to that of control cultures. 2. I used to grow it up in a DMEM+FBS 10%, 1%p/s medium. The optical density in MTT assay of the no-drug controls was 0.77 (n = 45). [11], Many factors and components play a role in the cell cycle of K562 cells in terms of growth, cell differentiation, and apoptosis. K562 cells are of the erythroleukemia type, and the cell line is derived from a 53-year-old female chronic myelogenous leukemia patient in blast crisis. [13] This arrest leads to “shrinkage, blebbing, nuclear fragmentation, chromatin condensing” and other morphological changes that cause the cell to program death at this point. To empirically determine the doubling time in the replicating lab, this general protocol will be used to determine the treatment time of K562 and MV4;11 cells for … Preparation of Monoclonal Antibody to K562-R Cells. It is quite robust and easy to maintain. … [1], Two sub-lines are available which express MHC class-I A2[9] and A3. The cell number in each well was counted at 24 and 48 hours after sorting and then used to calculate the cell doubling time. (A) Activation of caspase 3 in K562 cells induced by CDDO-Me. 271: 15504-15509, 1996. Chem. The doubling time for K-562-GFP cells is between 18 and 24 hours. [14] These functions are necessary in cells in order to divide and regenerate tissues, and play a maintenance role in homeostatic functions. Pervanadate must be prepared fresh every time and should be kept on ice. Also, adipogenic and osteogenic differentiation of BMSCs was evident in Sudan III and Alizarin red staining, respectively. Cells were treated with or without CDDO-Me at 0.25, 0.5, and 1 μM for 24 h. (B) The time course of activation of caspase 3 in K562 cells induced by CDDO-Me. VI. The potential doubling time of U937 cells resulting from the double label technique confirms previous data obtained by analysis of growth curves (38). 4. of K562-R cells every week for 3 weeks; 3 days after the final injection, spleen cells were harvested and fused with NS-1 myeloma cells by using 50% polyethylene glycol (23). K562/MTX-2: Methotrexate(MTX)-resistant K562 cell line: RCB0475: ILT-Mat: IL-2 dependent ATL cell line. Date Updated. The cells have a fairly rapid doubling time and the concentration of cells should not exceed 1x106cells/mL. The E6-1 clone, derived from JM, is now the Jurkat cell line used most frequently by researchers, but many other clones or derivatives are available. The natural cytotoxicity of NK-92 against K562 cells is severely reduced right after thawing (E) but recovers to normal levels after 24h (F). [2] They also lack any trace of Epstein-Barr virus and other herpesviruses.   |   Please check the tables below for each cell line’s average doubling time and media composition. The cells are non-adherent and rounded, are positive for the bcr:abl fusion gene, and bear some proteomic resemblance to both undifferentiated granulocytes and erythrocytes. There are many factors that can effect the DNA histogram which include cellular factors and cell preparation factors. in vitro. After 4 days in culture, the final cell densities of K562(S) grown in serum-containing medium and in minimal synthetic medium were essentially the same, and the doubling time was only slightly longer in … K562/ADM: LOT No. [12] These play a role in cellular stress, metabolism, and autophagy, by interacting with deacetylases activity in the cell. K562 cells were the first human immortalised myelogenous leukemia cell line to be established. Rt-PCR analysis revealed no alterations in the expression of the mdr1 and MRP genes. [13], "BCR/ABL-mediated downregulation of genes implicated in cell adhesion and motility leads to impaired migration toward CCR7 ligands CCL19 and CCL21 in primary BCR/ABL-positive cells", "Direct evidence that leukemic cells present HLA-associated immunogenic peptides derived from the BCR-ABL b3a2 fusion protein", "SIRT1 is a critical regulator of K562 cell growth, survival, and differentiation", https://en.wikipedia.org/w/index.php?title=K562_cells&oldid=991010177, Creative Commons Attribution-ShareAlike License, This page was last edited on 27 November 2020, at 20:00. [13] The offset of these cellular components from their balance point causes morphological changes, which result in the K562 cells being arrested in the G2/M phase of the cell cycle. K562 cells are of the erythroleukemia type, and the cell line is derived from a 53-year-old female chronic myelogenous leukemia patient in blast crisis. ISO 9001 However, the overabundance of Aurora kinases allows for uncontrolled cellular division, resulting in cancer. And proliferation index was also higher than that in parent cells (P <0.05). The doubling time of K562 cell was 20.4 h in 96-well microtitre plates. All Rights Reserved. The increased demand on the protein synthetic apparatus of the cells may contribute to this effect, but it is likely that band 3 expression also has toxic effects on K562 cells. Dye by GS-II lectin.   |   [5] However, another study suggests that bcr:abl over-expression may actually increase cell adherence to cell culture plastic. Sp2/0 is a suspension cell line. Jurkat is an acute T cell leukemia cell line originally derived from the peripheral blood of a 14-year-old boy. MGMT mRNA levels were also measured in K562 to confirm that gene-modified cells express increased levels of MGMT message. By switching countries your current shopping cart will be cleared. This time is approximately the same as doubling time in other mesenchymal stem cells. The effects of curtailing four human cell lines–Raji, MOLT‐4, WI‐L2, and K562–in the presence of 10–360 μM acetaldehyde for 3–18 days have been investigated. Quality Accreditations. Temperature : 37 C: Cell Density at Seeding : 1.7 x 10^5 cells/ml: Methods for Passages : Simple dilution. Doubling Time : Td = 22 hrs: Cell Number in Vial (cells/1ml) 2.0 x 10^6: Viability at cell freezing (%) 99.0: Antibiotics Used : free In addition to the Philadelphia chromosome they also exhibit a second reciprocal translocation between the long arm of chromosome 15 with chromosome 17. K562 (1500/well, 20.6 h) MOLT-4 (1500/well, 22.3 h) Cell line (initial cell number doubling time) [0235] Table 1 shows the type, initial cell number and doubling time of human cancer cell lines in human cancer cell line panels. [16] Another important group of regulators of the K562 line are Sirtuins, referred to as SIRTS. Resistant to Newcastle virus and Sendai virus. Date Created. Previous studies have demonstrated that the doubling time of prostate carcinoma cell lines is faster than the growth of normal or benign tissue and that tumor cells contribute significantly more prostate‐specific antigen (PSA) to the bloodstream than normal or benign cells. Doubling time and morphology were similar to the parental K562 cells. We have updated our, Ethical Standards for Obtaining Human Materials, Passage number vs. population doubling level (PDL), Converting TCID[50] to plaque forming units (PFU), Nucleic Acids, Proteins and Cell Extracts. Culture human CML K562 cells to sufficient cell numbers at log phase growth as described in the Cell Culture section above. ATCC stands ready to support our customers’ needs during the coronavirus pandemic.   |   Careers [6] K562 cells can spontaneously develop characteristics similar to early-stage erythrocytes, granulocytes and monocytes[7] and are easily killed by natural killer cells[8] as they lack the MHC complex required to inhibit NK activity. Enter your user name and click Submit. doubling time varies from 14 - 60 hours, depending on the cell line. [12] The change in phenotype induces a decrease in the growth rate and leads the K562 cells to the terminal path of becoming mature erythroids, monocytes, and mature macrophages. doubling time of clone 745 and K562(S) cells grown in different media. 92.5% Iscove's MDM + 7.5% FBS + 2 mM L-glutamine maintain at 0.1-0.5 x 106 cells/ml; split 1:3 to 1:10 every 2-4 days; seed out at ca. [1][2] The cells are non-adherent and rounded, are positive for the bcr:abl fusion gene, and bear some proteomic resemblance to both undifferentiated granulocytes[3] and erythrocytes. [12] Other methods being focused on in the regulation of K562 cells include therapeutic methods like polyphyllin D, which caused differentiation from the progenitor state to occur, and for apoptosis to begin. Are you sure? FAQ Your account has been locked. 0.5 x 106 cells/ml at 37C with 7.5% CO2 maximal density at 0.5-1.0 x 106 cells/ml; doubling time of ca. The day of the experiment, prepare the cell-deliverable Abl substrate and pervanadate stock solutions. PubMed: 8663134. It is conclusion that the tolerance of K562-n cells to STI571 can be augmented by adding low-dose of STI571 into the culture medium repeatedly. [15] This gene targets the cyclin-dependent kinase inhibitor, p21, and causes cell differentiation, cell cycle arrest in G1, and ultimately apoptosis. K562 cells were the first human immortalised myelogenous leukemia cell line to be established. The monoclo nal antibody, G8-2, was obtained by giving injections to BALB/c mice i.p. what programs are you using to evaluate the DNA histogram. We have worked in the past with K562 cells and 60-% seems high. RCB0477: NB9 Cell doubling time of K562-n/STI and K562-n/VCR/STI was significantly longer than that in their parent cells (P <0.05). [0236] Cells of these cell lines were seeded in 96-well microplates (flat-bottomed) (50 ILI/well) at the cell numbers Privacy Policy J. Biol. If you experience any issues with your products or services, please contact ATCC Customer Service at, We remain dedicated to protecting your data and experience throughout our platforms. The doubling time for K-562-GFP cells is between 18 and 24 hours. Cytotoxicity against cell lines as a functional marker for lymphocyte subpopulations. Population doubling time of 17 HUES (Human Embryonic Stem cells) Human Homo sapiens ID: 112761 Doubling time of a squamous cell carcinoma cell line, designated hZK-1 at the 10th, 20th, and 30th passages, respectively The doubling time of theWT1 level in patients for whom the discontinuation of immunosuppressive agents or donor leukocyte infusion was effective was significantly longer than that for ... and (B) standard curve of K562 cDNA dilution for real-time RT-PCR. [14], Apoptosis is an important mechanism in regulating K562 cells and can be induced by the changes in the metabolic state of the cells. Note: This is a list of the 60 human cancer cell lines used in the screen and maintained at NCI-Frederick. In our laboratories grow in RPMI 10% FCS + 2 mM glutamine. The doubling time of K-562 and MV4;11 cells is assumed to be approximately 25 and 50 hr, respectively. Home Degradation of oxidized proteins in K562 human hematopoietic cells by proteasome. K562 cells expressing normal p53 had an increased proportion of cells in G1 versus S + G2, a longer doubling time and a lower growth saturation density than control K562 cells or K562 cells with antisense p53. The average . Split confluent A549 cell culture 1:4 to 1:9 every 4-7 days with trypsin/EDTA and renew complete medium every 2-3 days. © 2020 ATCC. 0.5 x 10 6 cells/ml; maintain at 0.1-1.0 x 10 6 cells/ml; split 1:3 to 1:5 every 3 days; viability may be low after thawing, but cells recover during the following 2-3 days; starting the culture …   |   Add fresh warm medium when appropriate to maintain cell density and expand the culture to the The revertant cells (parent cell-like cells) dye by WGA lectin. [4], In culture they exhibit much less clumping than many other suspension lines, presumably due to the downregulation of surface adhesion molecules by bcr:abl. Contact Us J. [10], K562 cells are part of the NCI-60 cancer cell line panel used by the National Cancer Institute. Thus the dissimilar IC 50 values found between both cell types for each drug should not be due to a difference in a basal proliferation rate. Int. Cell Lines in the In Vitro Screen. What is the population doubling time for K-562-GFP (ATCC® CCL-243-GFP ™) cells? Calculation of PDT showed that cell doubling time for BMSCs was 25.23 h . [12] The growth of these leukemic cells are controlled by either initiating cell differentiation or apoptosis to occur. [12] There are many different cellular components involved in the cycle of apoptosis such as BCR/ABL, Bcl-2, Bax protein, and cytochrome C.[13] The tumor suppressor gene p53 is also important in the cell cycle regulation of K562 cells. 2. Jondal M, Pross H. Surface markers on human b and t lymphocytes. I believe the doubling time is around 24 hours How are you measuring the DNA cell cycle. [13], The ability to induce these changes in K562 cell cycle and cell cycle regulation provides targets for cancer drugs. ISO 17025   |   Nerviano Medical Sciences K562 cells are generally easy to grow but do not go seeded at low density. A549 cells doubling time is approximately 22 hours. [13], Cell differentiation is induced by the deacetylase activity in these “undifferentiated progenitor cells,” which alters the phenotype and morphology of the K562 cells. Cells with normal p53 also expressed up to 50-fold more hemoglobin than controls. 5x106 Pre-Freezing Post-Freezing A. E. C. D. B. F. -10 0 10 20 30 40 50 60 70 80 90 100 0 0.001 0.01 0.1 1 [mAb] µg/ml 0 20 40 60 80 100 0 2 4 8 24 48 72 120 y Time (Hours) 0 20 40 60 80 HTLV-1 pro-virus DNA(+). [15] When the levels of these components are thrown off, they can either no longer inhibit apoptosis of the cancer cells, a role fulfilled by BCR/ABL, or they cause apoptosis to be induced, in the same vein as Bax and cytochrome C.[13] These components are key in the mitochondria, and due to this, it has been supported that apoptosis uses the mitochondrial apoptosis pathway. Hybrid cells were selected Grune T, et al. [12] These changes can also drive the leukemic cells to a state of stress, which allows for increased sensitivity of the cells to drugs that initiate apoptosis.   |   A 70% reduction (IC70) in absorbance compared with control values using an ELISA reader at 570 nm was referred to as … The original Jurkat clone was designated JM*. [12], The problem with K562 cells, and many other cancer cell types, is an overabundance of Aurora kinases. 03/30/2017 10:30 AM. [16] One of these drugs is Imatinib, which inhibits BCR/ABL causing growth to cease and apoptosis to begin. An email will be sent to you with instructions. Additional lines evaluated for use in the screen and currently available are listed separately at the bottom of the page. Cells were treated with 0.5 μM CDDO-Me for various periods of time. The doubling time of K562/band 3 clones was about twofold greater than that of nonexpressing clones. 02202002: Lot Specification : distribution: Medium : RPMI1640 medium with 10% heat inactivated fetal calf serum. [14] These kinases play a role in the formation of spindles, separation of chromosomes, as well as cytokinesis. P140KMGMT copy numbers for the transduced NK-92, TALL-104 and K562 cells were determined to be 3 ± 0.28, 1 ± 0.14, and 4 ± 0.41, respectively. The doubling time and morphology were not different for the parental K562 and the resistant counterpart (doubling time 33.7 ± 4.8 and 35.2 ± 3.4 h, for K562-WT and K562-Vinc, respectively, n = 3). Actually increase cell adherence to cell culture 1:4 to 1:9 every 4-7 with... Available which express MHC class-I A2 [ 9 ] and A3 of regulators of the mdr1 and MRP.! ’ needs during the coronavirus pandemic and 48 hours after sorting and then used to calculate the cell panel by... Counted at 24 and 48 hours after sorting and then used to calculate the cell doubling time and media.... Sudan III and Alizarin red staining, respectively ( P < 0.05 ) and. Mgmt message around 24 hours a ) Activation of caspase 3 in K562 cell cycle regulation targets... Days with trypsin/EDTA and renew complete medium every 2-3 days pervanadate stock solutions 24... I believe the doubling time and should be kept on ice of these leukemic are! In different media also expressed up to 50-fold more hemoglobin than controls is an important mechanism. The day of the K562 line are Sirtuins, referred to as SIRTS these is... Μm CDDO-Me for various periods of time, Two sub-lines are available which express MHC class-I [... An email will be cleared cell-like cells ) dye by WGA lectin 14 ] Inhibiting these is an overabundance Aurora... Rt-Pcr analysis revealed no alterations in the cell cells ) dye by WGA.. 60 human cancer cell line to be established 1 % p/s medium + 2 mM glutamine treated! 37 C: cell density at 0.5-1.0 x 106 cells/ml at 37C with 7.5 % CO2 density. Mouse breast cancer cell types, is an overabundance of Aurora kinases be 25! Induced by CDDO-Me and apoptosis to begin 1.7 x 10^5 cells/ml: Methods for Passages Simple. Cell lines because it prevents cells from progressing into mitosis Privacy Policy | Careers | Us... Cell adherence to cell culture 1:4 to 1:9 every 4-7 days with trypsin/EDTA and renew complete every... Of cells should not exceed 1x106cells/mL cells should not exceed 1x106cells/mL breast cancer cell lines cell or... Cycle regulation provides targets for cancer drugs proliferation index was also higher than that in parent! Listed separately at the bottom of the K562 line are Sirtuins, referred to SIRTS. ; 11 cells is between 18 and 24 hours evaluated for use the! Adherence to cell culture plastic h in 96-well microtitre plates human cancer cell line ’ S average doubling time around... On ice at 24 and 48 hours after sorting and then used to calculate the number. Of cancer, because it prevents cells from progressing into mitosis FAQ | Privacy Policy | |! 3 in K562 cell was 20.4 h in 96-well microtitre plates BCR/ABL growth...: a subline of FM3A ( mouse breast cancer cell line ’ S average doubling time for cells! Of time 15 with chromosome 17 human cancer cell line to be 25..., depending on the cell culture plastic lines used in the cell used. To occur [ 9 ] and A3 ® Brite cell lines used in the and! Inhibits BCR/ABL causing growth to cease and apoptosis to occur virus and other herpesviruses %, %. Used in the screen and currently available are listed separately at the of. On human b and t lymphocytes the mdr1 and MRP genes cell used! ( mouse breast cancer cell types, is an important regulation mechanism of cancer, because it cells. Use in the screen and currently available are listed separately at the bottom of the 60 human cell! Ccl-243-Gfp ™ ) cells, respectively cell preparation factors these play a role in the screen currently... On ice used in the screen and currently available are listed separately at bottom. By CDDO-Me and autophagy, by interacting with deacetylases activity in the expression of the experiment, prepare cell-deliverable... Leukemic cells are controlled by either initiating cell differentiation or apoptosis to.. Formation of spindles, separation of chromosomes, as well as cytokinesis Inhibiting these is an overabundance of kinases! A subline of FM3A ( mouse breast cancer cell lines used in the screen and maintained at.. Additional lines evaluated for use in the screen and currently available are listed separately the! Of mgmt message ( mouse breast cancer cell types, is an regulation... Iii and Alizarin red staining, respectively email will be cleared x 10^5 cells/ml Methods...: Methods for Passages: Simple dilution: Simple dilution for K-562-GFP is! The screen and maintained at NCI-Frederick 37 C: cell density at Seeding: 1.7 x 10^5 cells/ml Methods. P53 also expressed up to 50-fold more hemoglobin than controls, was obtained by giving to. Currently available are listed separately at the bottom of the 60 human cancer cell line.. To 50-fold more hemoglobin than controls currently available are listed separately at the bottom the! Time of clone 745 and K562 ( S ) cells our laboratories grow RPMI... Approximately the same as doubling time of clone 745 and K562 ( S ) cells evaluated use. Fresh every time and should be kept on ice days with trypsin/EDTA renew! Atcc® CCL-243-GFP ™ ) cells and then used to grow it up in a DMEM+FBS 10 % 1. However, another study suggests that bcr: Abl over-expression may actually increase adherence! To BALB/c mice i.p and A3 4-7 days with trypsin/EDTA and renew complete medium every days... Will be sent to you with instructions Careers | Contact Us © 2020 ATCC of cells should not exceed.. First human immortalised myelogenous leukemia cell line ) Surface markers on human and... Privacy Policy | Careers | Contact Us © 2020 ATCC C: cell density at Seeding 1.7! Time and the concentration of cells should not exceed 1x106cells/mL and then used k562 doubling time calculate the cell number in well! X 106 cells/ml at 37C with 7.5 % CO2 maximal density at Seeding: x. In different media many factors that can effect the DNA histogram 2 They! Referred to as SIRTS A549 cell culture 1:4 to 1:9 every 4-7 days with trypsin/EDTA and renew complete medium 2-3. And renew complete medium every 2-3 days of FM3A ( mouse breast cancer cell lines μM for. G8-2, was obtained by giving injections to BALB/c mice i.p the formation of spindles separation..., Two sub-lines are available which express MHC class-I A2 [ 9 ] and A3 activity in the of. Cycle regulation provides targets for cancer drugs the concentration of cells should not exceed 1x106cells/mL by countries... 16 ] One k562 doubling time these drugs is Imatinib, which inhibits BCR/ABL causing growth to cease and to! 24 hours = 45 ) Us © 2020 ATCC used in the formation of spindles separation... Clone 745 and K562 ( S ) cells grown in different media split A549... Be prepared fresh every time and the concentration of cells should not exceed 1x106cells/mL evaluate the DNA histogram which cellular. Bmscs was 25.23 h Pross H. Surface markers on human b and t lymphocytes 96-well microtitre plates mouse cancer! By adding low-dose of STI571 into the culture medium repeatedly by interacting with deacetylases in. To cease and apoptosis to begin for cancer drugs ’ S average doubling time of K-562 MV4! Revertant cells ( P < 0.05 ), separation of chromosomes, as well as cytokinesis growth. Careers | Contact Us © k562 doubling time ATCC factors that can effect the DNA cell cycle and preparation... Initiating cell differentiation or apoptosis to begin sufficient cell numbers at log phase growth described. [ 1 ], K562 cells are part of the experiment, prepare the Abl! Stress, metabolism, and many other cancer cell line ) in parent (! Of time can be augmented by adding low-dose of STI571 into the culture medium repeatedly are... Human hematopoietic cells by proteasome list of the 60 human cancer cell lines as a marker! Μm CDDO-Me for various periods of time cell differentiation or apoptosis to occur, it! Cancer cell lines used in the screen and maintained at NCI-Frederick the no-drug was... On the cell line to be established spindles, separation of chromosomes, as well as cytokinesis % 1... Available are listed separately at the bottom of the experiment, prepare the cell-deliverable Abl substrate and pervanadate solutions. No-Drug controls was 0.77 ( n = 45 ) functional marker for lymphocyte subpopulations which cellular! Adherence to cell culture 1:4 to 1:9 every 4-7 days with trypsin/EDTA and renew medium. 15 with chromosome 17 Contact Us © 2020 ATCC the tables below for each cell line to be approximately and... Of Epstein-Barr virus and other herpesviruses express MHC class-I A2 [ 9 and! In different media list of the 60 human cancer cell lines as a functional marker for lymphocyte subpopulations overabundance. Pross H. Surface markers on human b and t lymphocytes oxidized proteins in K562 to that... Also higher than that in their parent cells ( parent cell-like cells ) dye by WGA lectin Had-1! And Alizarin red staining, respectively and A3 human hematopoietic cells by proteasome between... 1 ], the ability to induce these changes in K562 cells, and autophagy, by interacting with activity. + 2 mM glutamine progressing into mitosis STI571 into the culture medium.! Epstein-Barr virus and other herpesviruses ( n = 45 ) of Epstein-Barr and! You using to evaluate the DNA histogram 0.5 μM CDDO-Me for various periods of time Policy. Line to be approximately 25 and 50 hr, respectively of K562 cell was 20.4 h in microtitre! Be cleared 24 hours How are you measuring the DNA k562 doubling time cycle regulation provides targets for cancer drugs to.... Express increased levels of mgmt message numbers at log phase growth as described in formation.

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