Megakaryoblastic leukemia 1 (MKL1),identified as part of the t(1;22) translocation specific to acute megakaryoblastic leukemia,is highly expressed in differentiated muscle cells and promotes muscle differentiation by activating serum response factor (SRF). Here we show that Mkl1 expression is up-regulated during murine megakaryocytic differentiation and that enforced overexpression of MKL1 enhances megakaryocytic differentiation. When the human erythroleukemia (HEL) cell line is induced to differentiate with 12-O-tetradecanoylphorbol 13-acetate,overexpression of MKL1 results in an increased number of megakaryocytes with a concurrent increase in ploidy. MKL1 overexpression also promotes megakaryocytic differentiation of primary human CD34(+) cells cultured in the presence of thrombopoietin. The effect of MKL1 is abrogated when SRF is knocked down,suggesting that MKL1 acts through SRF. Consistent with these findings in human cells,knockout of Mkl1 in mice leads to reduced platelet counts in peripheral blood,and reduced ploidy in bone marrow megakaryocytes. In conclusion,MKL1 promotes physiologic maturation of human and murine megakaryocytes.
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产品类型：
产品号#：
09500
09600
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04902
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04971
04901
产品名：
BIT 9500血清替代物
StemSpan™ SFEM
StemSpan™ SFEM
MegaCult™-C胶原蛋白和不含细胞因子的培养基
胶原蛋白溶液
MegaCult™-C培养基无细胞因子
双室载玻片试剂盒
MegaCult™-C cfu染色试剂盒
MegaCult™-C不含细胞因子完整试剂盒
MegaCult™-C细胞因子完整试剂盒
MegaCult™-C细胞因子培养基
Halene S et al. (SEP 2010)
Blood 116 11 1942--50
Serum response factor is an essential transcription factor in megakaryocytic maturation.
Serum response factor (Srf) is a MADS-box transcription factor that is critical for muscle differentiation. Its function in hematopoiesis has not yet been revealed. Mkl1,a cofactor of Srf,is part of the t(1;22) translocation in acute megakaryoblastic leukemia,and plays a critical role in megakaryopoiesis. To test the role of Srf in megakaryocyte development,we crossed Pf4-Cre mice,which express Cre recombinase in cells committed to the megakaryocytic lineage,to Srf(F/F) mice in which functional Srf is no longer expressed after Cre-mediated excision. Pf4-Cre/Srf(F/F) knockout (KO) mice are born with normal Mendelian frequency,but have significant macrothrombocytopenia with approximately 50% reduction in platelet count. In contrast,the BM has increased number and percentage of CD41(+) megakaryocytes (WT: 0.41% ± 0.06%; KO: 1.92% ± 0.12%) with significantly reduced ploidy. KO mice show significantly increased megakaryocyte progenitors in the BM by FACS analysis and CFU-Mk. Megakaryocytes lacking Srf have abnormal stress fiber and demarcation membrane formation,and platelets lacking Srf have abnormal actin distribution. In vitro and in vivo assays reveal platelet function defects in KO mice. Critical actin cytoskeletal genes are down-regulated in KO megakaryocytes. Thus,Srf is required for normal megakaryocyte maturation and platelet production partly because of regulation of cytoskeletal genes.
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产品类型：
产品号#：
09500
09600
09650
04971
04902
04901
04963
04962
产品名：
BIT 9500血清替代物
StemSpan™ SFEM
StemSpan™ SFEM
MegaCult™-C细胞因子完整试剂盒
胶原蛋白溶液
MegaCult™-C细胞因子培养基
双室载玻片试剂盒
MegaCult™-C cfu染色试剂盒
M. S. Tavangar et al. (may 2020)
Clinical and experimental dental research
Differential expression of drug resistance genes in CD146 positive dental pulp derived stem cells and CD146 negative fibroblasts.
INTRODUCTION The stem cell portion of the dental pulp derived cultures (DPSCs) showed a higher resistance to cytotoxic effect of restorative dental materials compared to pulpal fibroblasts (DPFs). Here,we aimed to compare the expression of some drug resistant genes between these cells. METHODS AND MATERIALS To separate DPSCs from DPFs,we used magnetic cell sorting technique based on CD146 expression. To assess the stem cell properties,the positive and negative portions underwent colony forming assays and were induced to be differentiated into the adipocytes,osteoblasts,hepatocytes,and neural cells. Cell surface antigen panels were checked using immune fluorescence and flow-cytometry techniques. The mRNA expression of 14 ABC transporters including ABCA2,ABCB1,ABCB11,ABCC1,ABCC2,ABCC3,ABCC4,ABCC5-2,ABCC5-4,ABCC5-13,ABCC6,ABCC10,ABCC11,and ABCG2 genes was assessed,using quantitative RT-PCR technique. RESULTS Only the CD146 positive portion could be differentiated into the desired fates,and they formed higher colonies (16.7 ± 3.32 vs. 1.7 ± 1.67,p {\textless} .001). The cell surface antigen panels were the same,except for CD146 and STRO-1 markers which were expressed only in the positive portion. Among the ABC transporter genes studied,the positive portion showed a higher expression (approximately two-fold) of ABCA2,ABCC5-13,and ABCC5-2 genes. CONCLUSION Dental pulp stem cells which can be separated from dental pulp fibroblasts based on CD146 expression,express higher levels of some drug resistance genes which probably accounts for their features of more resistance to cytotoxic effects of some dental materials. This needs to be more validated in future.
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产品类型：
产品号#：
05412
产品名：
MesenCult™ 脂肪分化试剂盒 (人)
V. Cesarini et al. (aug 2019)
Scientific reports 9 1 12206
Regulation of PDE5 expression in human aorta and thoracic aortic aneurysms.
Aneurysms and dissections affecting thoracic aorta are associated with smooth muscle cell (SMC) dysfunction. NO/cGMP signaling pathway in smooth muscle cells has been shown to be affected in sporadic thoracic aortic aneurysms. We analyzed the mRNA levels of PDE5,a cGMP-hydrolyzing enzyme highly expressed in aortic SMCs,that regulates arterious vascular tone by lowering cGMP levels. We found that aortic tissue obtained from Marfan,tricuspid and bicuspid thoracic aneurysms expressed lower levels of PDE5 mRNA compared to control aortas. In particular,we found that affected aortas showed lower levels of all the PDE5A isoforms,compared to control aortas. Transfection of vascular SMCs (VSMCs) with NOTCH3 activated domain (NICD3) induced the expression of PDE5A1 and A3 protein isoforms,but not that of the corresponding mRNAs. VSMC stimulation with GSNO,a nitric oxide analogue or with 8-br-cGMP,but not with 8-br-cAMP,up-regulated PDE5 and NOTCH-3 protein levels,indicating a negative feedback loop to protect the arterial wall from excessive relaxation. Finally,we found that PDE5 is expressed early during human aorta development,suggesting that if loss of function mutations of PDE5 occur,they might potentially affect aortic wall development.
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Dobo I et al. (DEC 1999)
Journal of hematotherapy & stem cell research 8 6 601--7
Endogenous erythroid and megakaryocytic colony formation in serum-free, cytokine-free collagen gels.
We studied the suitability of collagen-based semisolid medium for assay of endogenous erythroid colony formation performed in myeloproliferative disorders. Bone marrow (BM) mononuclear cells (MNC) from 103 patients suspected of having polycythemia vera (PV,76 patients) or essential thrombocythemia (ET,27 patients) were grown in collagen-based,serum-free,cytokine-free semisolid medium. Colony analysis at day 8 or 10 showed that this collagen assay is specific,as endogenous growth of erythroid colonies was never observed in cultures of 16 healthy donors and 6 chronic myelogenous leukemia (CML) patients. Endogenous erythroid colony formation was observed in 53.3% of patients suspected of PV,with only 15.4% of positive cultures for patients with 1 minor PV criterion and 72% (p = 0.009) of positive cultures for patients with textgreater or =2 minor or 1 major PV criterion. Similarly,endogenous growth of erythroid colonies was found in 44.4% of patients suspected of ET,with 31.6% of positive cultures for patients with 1 ET criterion versus 75% for patients with textgreater or =2 ET criteria. In addition,we found that in collagen gels,tests of erythropoietin (EPO) hypersensitivity in the presence of 0.01 or 0.05 U/ml of EPO and tests of endogenous colony-forming units-megakaryocyte (CFU-MK) formation cannot be used to detect PV or ET,as these tests were positive for,respectively,21.4% and 50% of healthy donors and 83% and 50% of CML patients. A retrospective analysis suggests that collagen assays are more sensitive than methylcellulose assays to assess endogenous growth of erythroid colonies. In summary,serum-free collagen-based colony assays are simple and reliable assays of endogenous growth of erythroid colonies in myeloproliferative diseases. They also appear to be more sensitive than methylcellulose-based assays.
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产品类型：
产品号#：
04850
04900
04901
04902
04960
04961
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04963
04970
04971
04974
产品名：
MegaCult™-C含脂培养基
MegaCult™-C培养基无细胞因子
MegaCult™-C细胞因子培养基
胶原蛋白溶液
MegaCult™-C胶原蛋白和不含细胞因子的培养基
MegaCult™-C胶原蛋白和细胞因子培养基
MegaCult™-C cfu染色试剂盒
双室载玻片试剂盒
MegaCult™-C不含细胞因子完整试剂盒
MegaCult™-C细胞因子完整试剂盒
MegaCult™-C胶原蛋白和脂质培养基
Ito CY et al. (JAN 2003)
Blood 101 2 517--23
Hematopoietic stem cell and progenitor defects in Sca-1/Ly-6A-null mice.
Despite its wide use as a marker for hematopoietic stem cells (HSCs),the function of stem cell antigen-1 (Sca-1) (also known as lymphocyte activation protein-6A [Ly-6A]) in hematopoiesis remains poorly defined. We have previously established that Sca-1(-/-) T cells develop normally,although they are hyperresponsive to antigen. Here,we report detailed analysis of hematopoiesis in Sca-1-deficient animals. The differentiation potential of Sca-1-null bone marrow was determined from examination of the most mature precursors (culture colony-forming units [CFU-Cs]) to less committed progenitors (spleen CFUs [CFU-Ss]) to long-term repopulating HSCs. Sca-1-null mice are mildly thrombocytopenic with a concomitant decrease in megakaryocytes and their precursors. Bone marrow cells derived from Sca-1(-/-) mice also have decreased multipotential granulocyte,erythroid,macrophage,and megakaryocyte CFU (GEMM-CFU) and CFU-S progenitor activity. Competitive repopulation assays demonstrated that Sca-1(-/-) HSCs are at a competitive disadvantage compared with wild-type HSCs. To further analyze the potential of Sca-1(-/-) HSCs,serial transplantations were performed. While secondary repopulations using wild-type bone marrow completely repopulated Sca-1(-/-) mice,Sca-1(-/-) bone marrow failed to rescue one third of lethally irradiated wild-type mice receiving secondary bone marrow transplants from irradiation-induced anemia and contributed poorly to the surviving transplant recipients. These data strongly suggest that Sca-1 is required for regulating HSC self-renewal and the development of committed progenitor cells,megakaryocytes,and platelets. Thus,our studies conclusively demonstrate that Sca-1,in addition to being a marker of HSCs,regulates the developmental program of HSCs and specific progenitor populations.
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产品类型：
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03434
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产品名：
MethoCult™GF M3434
MethoCult™GF M3434
MegaCult™-C胶原蛋白和不含细胞因子的培养基
胶原蛋白溶液
MegaCult™-C培养基无细胞因子
MegaCult™-C胶原蛋白和细胞因子培养基
MegaCult™-C细胞因子培养基
双室载玻片试剂盒
MegaCult™-C cfu染色试剂盒
MegaCult™-C不含细胞因子完整试剂盒
MegaCult™-C细胞因子完整试剂盒
Kimura Y et al. (APR 2004)
Proceedings of the National Academy of Sciences of the United States of America 101 16 6015--20
Targeted mutations of the juxtamembrane tyrosines in the Kit receptor tyrosine kinase selectively affect multiple cell lineages.
Loss-of-function mutations in the murine dominant white spotting/c-kit locus affect a diverse array of biological processes and cell lineages and cause a range of phenotypes,including severe anemia,defective pigmentation,sterility,mast cell deficits,a lack of interstitial cells of Cajal,spatial learning memory deficits,and defects in peripheral nerve regeneration. Here we show that tyrosine residues 567 and 569 in the juxtamembrane (Jx) domain of the murine Kit receptor tyrosine kinase are crucial for the function of Kit in melanogenesis and mast cell development,but are dispensable for the normal development of erythroid,interstitial cells of Cajal and germ cells. Furthermore,adult mice lacking both tyrosines exhibit splenomegaly,dysregulation of B-cell and megakaryocyte development,and enlarged stomachs. Analysis of signal transduction events induced by the mutant receptors after ligand stimulation indicates that Jx tyrosine mutations diminish receptor autophosphorylation and selectively attenuate activation of extracellular signal-regulated kinase/mitogen-activated protein kinases. Together,these observations demonstrate that the Jx domain of Kit plays a cell-type specific regulatory role in vivo and illustrate how engineered mutations in Kit can be used to understand the complex biological and molecular events that result from activating a receptor tyrosine kinase.
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产品类型：
产品号#：
03434
03444
04960
04902
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04961
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04963
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产品名：
MethoCult™GF M3434
MethoCult™GF M3434
MegaCult™-C胶原蛋白和不含细胞因子的培养基
胶原蛋白溶液
MegaCult™-C培养基无细胞因子
MegaCult™-C胶原蛋白和细胞因子培养基
MegaCult™-C细胞因子培养基
双室载玻片试剂盒
MegaCult™-C cfu染色试剂盒
MegaCult™-C不含细胞因子完整试剂盒
MegaCult™-C细胞因子完整试剂盒
Campard D et al. (MAY 2006)
Stem cells (Dayton,Ohio) 24 5 1302--14
Multilevel regulation of IL-6R by IL-6-sIL-6R fusion protein according to the primitiveness of peripheral blood-derived CD133+ cells.
Interleukin-6 (IL-6) and its soluble receptor (sIL-6R) are major factors for maintenance and expansion of hematopoietic stem cells (HSCs). Sensitivity of HSCs to IL-6 has been previously studied,in part by measuring the expression of IL-6R on the membrane (mIL-6R). Several studies have described the regulation of cell surface expression of IL-6R by several cytokines,but the role of glycoprotein 130 activation has not yet been investigated. In this study,CD133(+) cells were purified from adult peripheral blood and were precultured in the absence or presence of 5-fluorouracil (5-FU) for selection of quiescent HSCs. Cells were cultured with continuous or pulsed stimulations of an IL-6-sIL-6R fusion protein (hyperinterleukin-6 [HIL-6]) to 1) detect mIL-6R by flow cytometry,2) assess mIL-6R and sIL-6R RNAs by reverse transcription-polymerase chain reaction,3) measure sIL-6R in supernatants by enzyme-linked immunosorbent assay,4) analyze cell-cycle status,and 5) perform long-term culture-initiating cell assays. The level of mIL-6R(-) cells was preserved by 5-FU incubation. HIL-6 increased steady-state mIL-6R RNA and expression rate on HSCs,independently of treatment with 5-FU. Enhanced production of sIL-6R was observed with short pulses of HIL-6 on CD133(+) 5-FU-pretreated cells. This overproduction of sIL-6R was abrogated by tumor necrosis factor-alpha protease inhibitor-1,an inhibitor of a disintegrin and metalloprotease proteases,suggesting the shedding of mIL-6R. This phenomenon was mediated through the phosphatidylinositol-3'-kinase pathway and was involved in the maintenance of primitive HSCs. In conclusion,expression and production of IL-6R are tightly regulated and stage specific. We assume that sIL-6R produced by shedding should be involved in autocrine and paracrine loops in the HSC microenvironment.
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产品类型：
产品号#：
09600
09650
04960
04902
04900
04961
04901
04963
04962
04970
04971
产品名：
StemSpan™ SFEM
StemSpan™ SFEM
MegaCult™-C胶原蛋白和不含细胞因子的培养基
胶原蛋白溶液
MegaCult™-C培养基无细胞因子
MegaCult™-C胶原蛋白和细胞因子培养基
MegaCult™-C细胞因子培养基
双室载玻片试剂盒
MegaCult™-C cfu染色试剂盒
MegaCult™-C不含细胞因子完整试剂盒
MegaCult™-C细胞因子完整试剂盒
Gu T-l et al. (JUL 2007)
Blood 110 1 323--33
A novel fusion of RBM6 to CSF1R in acute megakaryoblastic leukemia.
Activated tyrosine kinases have been frequently implicated in the pathogenesis of cancer,including acute myeloid leukemia (AML),and are validated targets for therapeutic intervention with small-molecule kinase inhibitors. To identify novel activated tyrosine kinases in AML,we used a discovery platform consisting of immunoaffinity profiling coupled to mass spectrometry that identifies large numbers of tyrosine-phosphorylated proteins,including active kinases. This method revealed the presence of an activated colony-stimulating factor 1 receptor (CSF1R) kinase in the acute megakaryoblastic leukemia (AMKL) cell line MKPL-1. Further studies using siRNA and a small-molecule inhibitor showed that CSF1R is essential for the growth and survival of MKPL-1 cells. DNA sequence analysis of cDNA generated by 5'RACE from CSF1R coding sequences identified a novel fusion of the RNA binding motif 6 (RBM6) gene to CSF1R gene generated presumably by a t(3;5)(p21;q33) translocation. Expression of the RBM6-CSF1R fusion protein conferred interleukin-3 (IL-3)-independent growth in BaF3 cells,and induces a myeloid proliferative disease (MPD) with features of megakaryoblastic leukemia in a murine transplant model. These findings identify a novel potential therapeutic target in leukemogenesis,and demonstrate the utility of phosphoproteomic strategies for discovery of tyrosine kinase alleles.
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产品类型：
产品号#：
03434
03444
04960
04902
04900
04961
04901
04963
04962
04970
04971
产品名：
MethoCult™GF M3434
MethoCult™GF M3434
MegaCult™-C胶原蛋白和不含细胞因子的培养基
胶原蛋白溶液
MegaCult™-C培养基无细胞因子
MegaCult™-C胶原蛋白和细胞因子培养基
MegaCult™-C细胞因子培养基
双室载玻片试剂盒
MegaCult™-C cfu染色试剂盒
MegaCult™-C不含细胞因子完整试剂盒
MegaCult™-C细胞因子完整试剂盒
Dobo I et al. (AUG 1995)
Journal of hematotherapy 4 4 281--7
Collagen matrix: an attractive alternative to agar and methylcellulose for the culture of hematopoietic progenitors in autologous transplantation products.
Autografts using untreated or in vitro manipulated bone marrow and peripheral blood stem cells represent promising approaches to the treatment of malignant diseases. In this work,the collagen gel culture technique was compared with agar and methylcellulose for its capacity to permit the growth of human granulomonocytic (day 14 CFU-GM; collagen vs agar or MTC) or erythroblastic (day 7 CFU-E and day 14 BFU-E; collagen versus methylcellulose) colonies in autologous transplantation products. Our results show that the collagen culture system always gave as many or more colonies than the other techniques. It also allowed harvesting of gels onto glass slides and subsequent May-Grünwald-Giemsa,cytochemical or immunocytochemical staining. We suggest that the collagen assay represents an interesting alternative to the widely used agar or methylcellulose systems for the culture of hematopoietic progenitors because of the equal or higher number of colonies detected,the easy phenotypical identification of colonies in stained gels,and the ability to store high-quality documentation. This technique is particularly attractive for use in the quality control of autologous bone marrow transplantation procedures.
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产品类型：
产品号#：
04850
04900
04901
04902
04960
04961
04962
04963
04970
04971
04974
产品名：
MegaCult™-C含脂培养基
MegaCult™-C培养基无细胞因子
MegaCult™-C细胞因子培养基
胶原蛋白溶液
MegaCult™-C胶原蛋白和不含细胞因子的培养基
MegaCult™-C胶原蛋白和细胞因子培养基
MegaCult™-C cfu染色试剂盒
双室载玻片试剂盒
MegaCult™-C不含细胞因子完整试剂盒
MegaCult™-C细胞因子完整试剂盒
MegaCult™-C胶原蛋白和脂质培养基
Hogge D et al. (MAR 1997)
British journal of haematology 96 4 790--800
Quantitation and characterization of human megakaryocyte colony-forming cells using a standardized serum-free agarose assay.
Human progenitors of the megakaryocyte (Mk) lineage were detected by their ability to generate colonies-containing from 3 to textgreater 100 Mk,detectable as glycoprotein IIb/IIIa+ cells in APAAP-stained whole mount agarose cultures. Optimal growth conditions were achieved through the use of a defined serum substitute and a suitable cocktail of recombinant cytokines. Under these culture conditions,the smallest Mk-containing colonies (CFC-Mk) were detectable within a week followed by colonies containing larger numbers of Mk over the ensuing 2 weeks. The total number of CFC-Mk at 18-21 d was linearly related to the number of cells plated. Variation in the cytokines added showed that thrombopoietin (TPO) or IL-3 alone would support the formation of large numbers of CFC-Mk. However,optimal yields of colonies containing cells of both Mk and non-Mk lineages required the addition of other growth factors,of which a combination of IL-3,IL-6,GM-CSF and Steel factor (SF) +/- TPO was the best of those tested. The further addition of erythropoietin to this combination reduced the number of large pure' Mk colonies seen and in their place a corresponding number of mixed erythroid-Mk colonies became detectable. Flt3-ligand alone was unable to support the growth of CFC-Mk nor did it enhance their growth when combined with other factors. Plating of FACS-sorted sub-populations of CD34+ marrow cells in both serum-free agarose and methylcellulose assays demonstrated that most CFC-Mk are generated from CD34+ cells that are CD45RA- and CD71+�
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EasySep™小鼠TIL（CD45）正选试剂盒
科学海报Detection of Human Bipotential Erythroid-Megakaryocytic Progenitors in Serum-Free Collagen Gels
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