A human AML cell line (THP-1) was purchased from Chinese Academy of Sciences (Shanghai, China) and a human AML cell line transformed from MDS cells (SKM-1) was obtained from the Japanese Collection of Research Bioresources (LCRB; Tokyo, Japan). 293T cells were used as a control (Chinese Academy of Sciences, Shanghai, China). All cell lines were cultured in Roswell Park Memorial Institute-1640 medium (RPMI-1640; Hyclone; GE Healthcare Life Sciences) supplemented with 10% fetal bovine serum (Gibco-BRL; Thermo Fisher Scientific, Inc.) in a humidified incubator with 5% CO2 at 37°C.
Skm 1
Manufactured by Japanese Collection of Research Bioresources Cell Bank
Sourced in China, Japan
The SKM-1 is a piece of laboratory equipment designed for centrifugation. It is used to separate and concentrate cellular components, such as proteins, nucleic acids, and organelles, from complex biological samples. The SKM-1 can accommodate various rotor types and sample volumes, allowing it to be utilized in a wide range of research applications.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Mv4 11, by American Type Culture Collection (2 mentions)
Decitabine, by Merck Group (1 mentions)
Rpmi 1640, by GE Healthcare (1 mentions)
Deferasirox, by Novartis (1 mentions)
Thp 1, by American Type Culture Collection (1 mentions)
Kasumi 1, by Japanese Collection of Research Bioresources Cell Bank (1 mentions)
Molm 14, by Japanese Collection of Research Bioresources Cell Bank (1 mentions)
Nih3t3, by Japanese Collection of Research Bioresources Cell Bank (1 mentions)
Rpmi8226, by American Type Culture Collection (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Jurkat, by American Type Culture Collection (1 mentions)
Daudi, by American Type Culture Collection (1 mentions)
Molm 13, by Japanese Collection of Research Bioresources Cell Bank (1 mentions)
Mm 1s, by American Type Culture Collection (1 mentions)
Namalwa, by American Type Culture Collection (1 mentions)
Thp 1, by Japanese Collection of Research Bioresources Cell Bank (1 mentions)
Nomo 1, by Japanese Collection of Research Bioresources Cell Bank (1 mentions)
Karpas299, by European Collection of Authenticated Cell Cultures (1 mentions)
7 protocols using skm 1
1
Establishing AML and Control Cell Lines
2
Leukemia Cell Line Culturing Protocol
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The iron chelator Deferasirox was kindly donated by Novartis Pharma (Alcon (China) Ophthalmic Product, Shanghai, China) and the methylation drug Decitabine was purchased from Sigma (Sigma-Alorich, Shanghai, China). Human myeloid leukemia cell lines THP-1 and K562 were obtained from the Chinese Academy of Sciences, and SKM-1 from the Japanese Collection of Research Bioresources. All cell lines used in this study were cultured in RPMI 1640 (Hyclone; GE Healthcare Life Sciences, Logan UT, USA) supplemented with 10% fetal bovine serum (FBS; Gibco; Thermo Fisher Scienti c, Inc., Waltham, MA, USA), at 37°C with 5% CO2.
3
Cell Line Maintenance for Blood Cancer Research
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SKM-1 (MDS cell line), MOLM-14 and Kasumi-1 (AML cell lines), and NIH3T3 (Mouse fibroblast-like cell line) cells were purchased from the Japanese Collection of Research Bioresources Cell Bank (Ibaraki, Osaka, Japan). U937, THP-1, and MV4-11 (AML cell lines) cells and the human marrow stromal cell line HS-5 were purchased from the American Type Culture Collection (Manassas, VA, USA). MDSL (MDS cell line) cells were kindly provided to us by Professor Kaoru Tohyama (Kawasaki Medical School, Kurashiki City, Okayama, Japan). These cell lines were grown in Roswell Park Memorial Institute 1640 (RPMI 1640) medium, which contained 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin. MDS-L cells were cultured in RPMI 1640 medium with 20% FBS. NIH3T3 and HS-5 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) with 10% FBS.
4
Cell Line Panel for Hematological Cancers
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T cell non-Hodgkin’s lymphoma cell line KARPAS299 was supplied by the European Collection of Authenticated Cell Cultures (ECACC). Multiple myeloma cell lines, MM.1S and RPMI8226; Burkitt’s lymphoma cell lines, Daudi, Raji, and NAMALWA; the acute myelogenous leukemia cell line, KG1; the acute T cell leukemia cell line, Jurkat; and the chronic myelogenous leukemia cell line, K562, were supplied by American Type Culture Collection (ATCC). Acute monocytic leukemia cell lines, THP-1, MOLM-13, and NOMO-1; and the myelodysplastic syndrome cell line, SKM-1, were supplied by Japanese Collection of Research Bioresources Cell Bank (JCRB). All these cell lines were maintained in RPMI 1640 (Gibco BRL) supplemented with 10% heat-inactivated fetal bovine serum (Sigma), 100 µg/mL streptomycin, and 100 U/mL penicillin.
5
Cell Culture of SKM-1 MDS Cell Line
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SKM-1, an MDS cell line, was obtained from the Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan) (16 (link)). The cells were cultured in RPMI-1640 medium (Gibco Life Technologies, Carlsbad, CA, USA) containing 10% fetal bovine serum (Gibco Life Technologies), 100 U/ml penicillin (Sigma-Aldrich, St. Louis, MO, USA) and 100 µg/ml streptomycin (Sigma-Aldrich) in an environment of saturated humidity, 5% CO2 at 37°C. Cells in the logarithmic growth phase were used in all experiments.
6
Evaluation of AML Cell Lines and Compounds
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Human AML cell lines, including MV-4-11, MOLM-13, and HL-60, were purchased from American Type Culture Collection (ATCC; Manassas, VA, USA). Human AML cell line Ontario Cancer Institute‒Acute Myeloid Leukemia-3 (OCI-AML-3) was sourced from Cobioer (Nanjing, China) and SKM-1 from the Japanese Collection of Research Bioresources Cell Bank (JCRB). MV-4–11 cells were propagated in Iscove’s Modified Dulbecco’s Medium (IMDM, Gibco, Grand Island, NY, USA Cat# C12440500BT) supplemented with 10% fetal calf serum (FCS, AUSGENEX, Loganholme, QLD, Australia, Cat# FBSSA500-S). Other cell lines were propagated in Roswell Park Memorial Institute (RPMI) 1640 medium (Gibco, Cat# C11875500BT) containing 10% FCS. All experiments utilized genetically authenticated, microbial-free cells in their exponential phases of growth.
APG-115 (produced by Ascentage Pharma, Jiangsu, China) was dissolved in dimethyl sulfoxide (DMSO, Sigma, St. Louis, MO, USA, Cat# D8418) for in vitro experiments or suspended in 0.2% hydroxypropyl methylcellulose (HPMC, Sigma, Cat# H7509-25G) for in vivo animal studies. DAC (Cat# S120009), AZA (Cat# S178206), and Ara-C (Cat# S164806) purchased from Selleckchem were dissolved in DMSO for in vitro assays and saline for in vivo studies. RG-7388 (idasanutlin, MDM2-P53 inhibitor, MedChemExpress, Cat# HY-15676) was dissolved in DMSO for in vitro assays.
APG-115 (produced by Ascentage Pharma, Jiangsu, China) was dissolved in dimethyl sulfoxide (DMSO, Sigma, St. Louis, MO, USA, Cat# D8418) for in vitro experiments or suspended in 0.2% hydroxypropyl methylcellulose (HPMC, Sigma, Cat# H7509-25G) for in vivo animal studies. DAC (Cat# S120009), AZA (Cat# S178206), and Ara-C (Cat# S164806) purchased from Selleckchem were dissolved in DMSO for in vitro assays and saline for in vivo studies. RG-7388 (idasanutlin, MDM2-P53 inhibitor, MedChemExpress, Cat# HY-15676) was dissolved in DMSO for in vitro assays.
7
Characterizing Human AML and MDS Cell Lines
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Cell lines and cell culture conditions. Human AML cell lines, including HEL, U937 and THP-1, were propagated in a monolayer culture in RPMI-1640 medium. MDS cell line SKM-1 was purchased from the Japanese Collection of Research Bioresources. RPMI-1640 medium was supplemented with 15% fetal bovine serum, 100 U/ml penicillin, and 100 mg/ml streptomycin. The medium and antibiotics were bought from Invitrogen (Carlsbad, CA, USA). All cells were maintained in a 37˚C incubator with 95% humidity and 5% CO 2 .
Patient samples. Bone marrow samples were collected during routine diagnostic assessment after written informed consent had been obtained. The patients were diagnosed by using WHO classification. Patients' bone marrow MNCs were separated by Ficoll-Hypaque (Sigma Chemical Co.) densitygradient centrifugation and used immediately. All participants provided written informed consent prior to entering the study. The study was approved by the Institutional Review Board of the Affiliated Hospital of Guiyang Medical College.
Chemicals and antibodies. AZA (99.0% purity) was obtained from the Shanghai Huilun Life Science and Technology Corp. Antibodies for western blot analysis were obtained from Cell Signaling Technology (Beverly, MA, USA) and secondary antibodies were purchased from Li-Cor Corp. (Lincoln, NE, USA).
Patient samples. Bone marrow samples were collected during routine diagnostic assessment after written informed consent had been obtained. The patients were diagnosed by using WHO classification. Patients' bone marrow MNCs were separated by Ficoll-Hypaque (Sigma Chemical Co.) densitygradient centrifugation and used immediately. All participants provided written informed consent prior to entering the study. The study was approved by the Institutional Review Board of the Affiliated Hospital of Guiyang Medical College.
Chemicals and antibodies. AZA (99.0% purity) was obtained from the Shanghai Huilun Life Science and Technology Corp. Antibodies for western blot analysis were obtained from Cell Signaling Technology (Beverly, MA, USA) and secondary antibodies were purchased from Li-Cor Corp. (Lincoln, NE, USA).
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