Griffiths RE et al. (DEC 2007)
Blood 110 13 4518--25
Normal prion protein trafficking in cultured human erythroblasts.
Normal prion protein (PrP(c)),an essential substrate for development of prion disease,is widely distributed in hematopoietic cells. Recent evidence that variant Creutzfeldt-Jakob disease can be transmitted by transfusion of red cell preparations has highlighted the need for a greater understanding of the biology of PrP(c) in blood and blood-forming tissues. Here,we show that in contrast to another glycosylphosphoinositol-anchored protein CD59,PrP(c) at the cell surface of cultured human erythroblasts is rapidly internalized through the endosomal pathway,where it colocalizes with the tetraspanin CD63. In the plasma membrane,PrP(c) colocalizes with the tetraspanin CD81. Cross-linking with anti-PrP(c) or anti-CD81 causes clustering of PrP(c) and CD81,suggesting they can share the same microdomain. These data are consistent with a role for tetraspanin-enriched microdomains in trafficking of PrP(c). These results,when taken together with recent evidence that exosomes released from cells as a result of endosomal-mediated recycling to the plasma membrane contain prion infectivity,provide a pathway for the propagation of prion diseases.
View Publication
产品类型：
产品号#：
09600
09650
产品名：
StemSpan™ SFEM
StemSpan™ SFEM
Mandal PK and Rossi DJ (MAR 2013)
Nature protocols 8 3 568--82
Reprogramming human fibroblasts to pluripotency using modified mRNA
Induced pluripotent stem (iPS) cells hold the potential to revolutionize regenerative medicine through their capacity to generate cells of diverse lineages for future patient-specific cell-based therapies. To facilitate the transition of iPS cells to clinical practice,a variety of technologies have been developed for transgene-free pluripotency reprogramming. We recently reported efficient iPS cell generation from human fibroblasts using synthetic modified mRNAs. Here we describe a stepwise protocol for the generation of modified mRNA-derived iPS cells from primary human fibroblasts,focusing on the critical parameters including medium choice,quality control,and optimization steps needed for synthesizing modified mRNAs encoding reprogramming factors and introducing these into cells over the course of 2-3 weeks to ensure successful reprogramming. The protocol described herein is for reprogramming of human fibroblasts to pluripotency; however,the properties of modified mRNA make it a powerful platform for protein expression,which has broad applicability in directed differentiation,cell fate specification and therapeutic applications.
View Publication
产品类型：
产品号#：
05854
05855
05850
05857
05870
05875
36254
85850
85857
85870
85875
产品名：
mFreSR™
mFreSR™
DMEM/F-12 with 15 mM HEPES
mTeSR™1
mTeSR™1
N. J. Giridhar et al. (Sep 2025)
Biology Open 14 9
Temporal transcriptomic profiling of human three-dimensional neuromuscular co-cultures
The principal organization of mammalian neuromuscular junctions (NMJs) shares essential features across species. However,human NMJs (hNMJs) exhibit distinct structural and physiological properties. While recent advances in stem-cell-based systems have significantly improved in vitro modeling of hNMJs,the extent to which these models recapitulate in vivo development remains unclear. Here,we performed temporal transcriptomic analysis of human three-dimensional (3D) neuromuscular co-cultures,composed of iPSC-derived motoneurons and skeletal muscle engineered from primary myoblasts. We found that the expression pattern follows a temporally coordinated gene expression program underlying NMJ maturation. The model recapitulates transcriptional features of NMJ development,including early myoblast fusion and presynaptic development,followed by a late-stage upregulation of postsynaptic markers and embryonic AChR subunits. Importantly,comparable transcriptional dynamics across two independent hiPSC lines confirm the reproducibility and robustness of this system. This study confirms on a transcriptional level that human 3D neuromuscular co-cultures are a robust and physiologically relevant model for investigating hNMJ development and function.
View Publication
Goransson O et al. ( 2007)
Journal of Biological Chemistry 282 45 32549--32560
Mechanism of Action of A-769662, a Valuable Tool for Activation of AMP-activated Protein Kinase
We have studied the mechanism of A-769662,a new activator of AMP-activated protein kinase (AMPK). Unlike other pharmacological activators,it directly activates native rat AMPK by mimicking both effects of AMP,i.e. allosteric activation and inhibition of dephosphorylation. We found that it has no effect on the isolated alpha subunit kinase domain,with or without the associated autoinhibitory domain,or on interaction of glycogen with the beta subunit glycogen-binding domain. Although it mimics actions of AMP,it has no effect on binding of AMP to the isolated Bateman domains of the gamma subunit. The addition of A-769662 to mouse embryonic fibroblasts or primary mouse hepatocytes stimulates phosphorylation of acetyl-CoA carboxylase (ACC),effects that are completely abolished in AMPK-alpha1(-/-)alpha2(-/-) cells but not in TAK1(-/-) mouse embryonic fibroblasts. Phosphorylation of AMPK and ACC in response to A-769662 is also abolished in isolated mouse skeletal muscle lacking LKB1,a major upstream kinase for AMPK in this tissue. However,in HeLa cells,which lack LKB1 but express the alternate upstream kinase calmodulin-dependent protein kinase kinase-beta,phosphorylation of AMPK and ACC in response to A-769662 still occurs. These results show that in intact cells,the effects of A-769662 are independent of the upstream kinase utilized. We propose that this direct and specific AMPK activator will be a valuable experimental tool to understand the physiological roles of AMPK.
View Publication
产品类型：
产品号#：
72922
72924
产品名：
A769662
A769662, 50 mg
Regala RP et al. (OCT 2009)
Cancer research 69 19 7603--11
Atypical protein kinase Ciota is required for bronchioalveolar stem cell expansion and lung tumorigenesis.
Protein kinase Ciota (PKCiota) is an oncogene required for maintenance of the transformed phenotype of non-small cell lung cancer cells. However,the role of PKCiota in lung tumor development has not been investigated. To address this question,we established a mouse model in which oncogenic Kras(G12D) is activated by Cre-mediated recombination in the lung with or without simultaneous genetic loss of the mouse PKCiota gene,Prkci. Genetic loss of Prkci dramatically inhibits Kras-initiated hyperplasia and subsequent lung tumor formation in vivo. This effect correlates with a defect in the ability of Prkci-deficient bronchioalveolar stem cells to undergo Kras-mediated expansion and morphologic transformation in vitro and in vivo. Furthermore,the small molecule PKCiota inhibitor aurothiomalate inhibits Kras-mediated bronchioalveolar stem cell expansion and lung tumor growth in vivo. Thus,Prkci is required for oncogene-induced expansion and transformation of tumor-initiating lung stem cells. Furthermore,aurothiomalate is an effective antitumor agent that targets the tumor-initiating stem cell niche in vivo. These data have important implications for PKCiota as a therapeutic target and for the clinical use of aurothiomalate for lung cancer treatment.
View Publication
产品类型：
产品号#：
18555
18555RF
18554
18554RF
18564
18564RF
产品名：
Jani V et al. (NOV 2016)
Human immunology
Root cause analysis of limitations of virtual crossmatch for kidney allocation to highly-sensitized patients.
Efficient allocation of deceased donor organs depends upon effective prediction of immunologic compatibility based on donor HLA genotype and recipient alloantibody profile,referred to as virtual crossmatching (VCXM). VCXM has demonstrated utility in predicting compatibility,though there is reduced efficacy for patients highly sensitized against allogeneic HLA antigens. The recently revised deceased donor kidney allocation system (KAS) has increased transplantation for this group,but with an increased burden for histocompatibility testing and organ sharing. Given the limitations of VCXM,we hypothesized that increased organ offers for highly-sensitized patients could result in a concomitant increase in offers rejected due to unexpectedly positive crossmatch. Review of 645 crossmatches performed for deceased donor kidney transplantation at our center did not reveal a significant increase in positive crossmatches following KAS implementation. Positive crossmatches not predicted by VCXM were concentrated among highly-sensitized patients. Root cause analysis of VCXM failures identified technical limitations of anti-HLA antibody testing as the most significant contributor to VCXM error. Contributions of technical limitations including additive/synergistic antibody effects,prozone phenomenon,and antigens not represented in standard testing panels,were evaluated by retrospective testing. These data provide insight into the limitations of VCXM,particularly those affecting allocation of kidneys to highly-sensitized patients.
View Publication
产品类型：
产品号#：
15223
15263
产品名：
RosetteSep™人总淋巴细胞富集抗体混合物
RosetteSep™人总淋巴细胞富集抗体混合物
Ting S et al. (MAY 2014)
Biotechnology journal 9 5 675--683
Time-resolved video analysis and management system for monitoring cardiomyocyte differentiation processes and toxicology assays.
Cardiomyocytes (CM) derived from human embryonic stem cells (hESC) are used for cardio-toxicity evaluation and tested in many preclinical trials for their potential use in regenerative therapeutics. As more efficient CM differentiation protocols are developed,reliable automated platforms for characterization and detection are needed. An automated time-resolved video analysis and management system (TVAMS) has been developed for the evaluation of hESC differentiation to CM. The system was used for monitoring the kinetics of embryoid bodies (EB) generation (numbers and size) and differentiation into beating EBs (percentage beating area and beating EB count) in two differentiation protocols. We show that the percentage beating areas of EBs (from total area of the EBs) is a more sensitive and better predictor of CM differentiation efficiency than percentage of beating EBs (from total EBs) as the percentage beating areas of EBs correlates with cardiac troponin-T and myosin heavy chain expression levels. TVAMS can also be used to evaluate the effect of drugs and inhibitors (e.g. isoproterenol and ZD7288) on CM beating frequency. TVAMS can reliably replace the commonly practiced,time consuming,manual counting of total and beating EBs during CM differentiation. TVAMS is a high-throughput non-invasive video imaging platform that can be applied for the development of new CM differentiation protocols,as well as a tool to conduct CM toxicology assays.
View Publication
产品类型：
产品号#：
05850
05857
05870
05875
85850
85857
85870
85875
产品名：
mTeSR™1
mTeSR™1
S. Mishra et al. (apr 2022)
Bio-protocol 12 8 e4391
An Optimized Tat/Rev Induced Limiting Dilution Assay for the Characterization of HIV-1 Latent Reservoirs.
The administration of antiretroviral therapy (ART) leads to a rapid reduction in plasma viral load in HIV-1 seropositive subjects. However,when ART is suspended,the virus rebounds due to the presence of a latent viral reservoir. Several techniques have been developed to characterize this latent viral reservoir. Of the various assay formats available presently,the Tat/Rev induced limiting dilution assay (TILDA) offers the most robust and technically simple assay strategy. The TILDA formats reported thus far are limited by being selective to one or a few HIV-1 genetic subtypes,thus,restricting them from a broader level application. The novel TILDA,labelled as U-TILDA ('U' for universal),can detect all the major genetic subtypes of HIV-1 unbiasedly,and with comparable sensitivity of detection. U-TILDA is well suited to characterize the latent reservoirs of HIV-1 and aid in the formulation of cure strategies. Graphical abstract.
View Publication
EasySep™小鼠TIL（CD45）正选试剂盒
技术公告Tumorsphere Culture of Human Breast Cancer Cell Lines
新闻STEMCELL Technologies Signs Exclusive License Agreement with Cincinnati Children’s for Stem Cell-Derived Organoid Technology
沪公网安备31010102008431号