技术资料

Ng WL et al. (JAN 2014) Cell death & disease 5 1 e1024

OCT4 as a target of miR-34a stimulates p63 but inhibits p53 to promote human cell transformation

Human cell transformation is a key step for oncogenic development,which involves multiple pathways; however,the mechanism remains unclear. To test our hypothesis whether cell oncogenic transformation shares some mechanisms with the process of reprogramming non-stem cells to induced pluripotent stem cells (iPSC),we studied the relationship among the key factors for promoting or inhibiting iPSC in radiation-transformed human epithelial cell lines derived from different tissues (lung,breast and colon). We unexpectedly found that p63 and OCT4 were highly expressed (accompanied by low expressed p53 and miR-34a) in all transformed cell lines examined when compared with their non-transformed counterparts. We further elucidated the relationship of these factors: the 3p strand of miR-34a directly targeted OCT4 by binding to the 3′ untranslated region (3′-UTR) of OCT4 and,OCT4,in turn,stimulated p63 but inhibited p53 expression by binding to a specific region of the p63 or p53 promoter. Moreover,we revealed that the effects of OCT4 on promoting cell oncogenic transformation were by affecting p63 and p53. These results support that a positive loop exists in human cells: OCT4 upregulation as a consequence of inhibition of miR-34a,promotes p63 but suppresses p53 expression,which further stimulates OCT4 upregulation by downregulating miR-34a. This functional loop contributes significantly to cell transformation and,most likely,also to the iPSC process. View Publication

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05850

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60062

60062AD

60062AD.1

60062BT

60062FI

60062FI.1

60062PE

60062PE.1

60060

60060AD

60060AD.1

60060BT

60060FI

60060FI.1

60060PE

60060PE.1

85850

85857

85870

85875

产品名：

抗人SSEA-4抗体，克隆号MC-813-70，生物素

抗人SSEA-4抗体，克隆号MC-813-70，FITC

抗人SSEA-4抗体, 克隆号MC-813-70，FITC

抗人SSEA-4抗体，克隆号MC-813-70，PE

抗小鼠SSEA-1（CD15）抗体，克隆MC-480

抗小鼠SSEA-1（CD15）抗体，克隆MC-480，Alexa Fluor® 488

抗小鼠SSEA-1（CD15）抗体，克隆MC-480，Biotin

抗小鼠SSEA-1（CD15）抗体，clone MC-480，FITC

抗小鼠SSEA-1（CD15）抗体，克隆MC-480，FITC

抗小鼠SSEA-1（CD15）抗体，克隆MC-480，PE

mTeSR™1

Hanson V et al. (OCT 2013) Tissue antigens 82 4 269--75

Assessment of the purity of isolated cell populations for lineage-specific chimerism monitoring post haematopoietic stem cell transplantation.

Following haematopoietic stem cell transplantation,monitoring the proportion of donor and recipient haematopoiesis in the patient (chimerism) is an influential tool in directing further treatment choices. Short tandem repeat (STR) analysis is a method of chimerism monitoring using DNA isolated from peripheral blood,bone marrow or specific isolated cell lineages such as CD3+ T cells. For lineage-specific STR analysis on cell populations isolated from peripheral blood,a qualitative estimation of the purity of each isolated population is essential for the correct interpretation of the test data. We describe a rapid,inexpensive method for the determination of purity using a simple flow cytometry method. The method described for assessing the purity of sorted CD3+ cells can be applied to any cell population isolated using the same technology. Data obtained were comparable to results from a commercial polymerase chain reaction (PCR)-based method for the assessment of purity (Non-T Genomic Detection Kit,Accumol,Calgary,AB,Canada) (P = 0.59). Of the 303 samples tested by flow cytometry,290 (95.7%) exceeded 90% purity,and 215 (70.95%) were over 99% pure. There were some outlying samples,showing diversity between samples and the unpredictability of purity of isolated cell populations. This flow cytometry method can be easily assimilated into routine testing protocols,allowing purity assessment in multiple-sorted cell populations for lineage-specific chimerism monitoring using a single secondary antibody and giving results comparable to a PCR-based method. As purity of isolated cell lineages is affected by time after venepuncture and storage temperature,assessment of each sample is recommended to give a reliable indication of sample quality and confidence in the interpretation of the results. View Publication

产品号#：

21000

20119

20155

18081

18081RF

产品名：

RoboSep™- S

RoboSep™ 吸头组件抛光剂

RoboSep™分选管套装（9个塑料管）

Herrera VL et al. ( 2014) PloS one 9 1 e85821

DEspR roles in tumor vasculo-angiogenesis, invasiveness, CSC-survival and anoikis resistance: a 'common receptor coordinator' paradigm.

A priori,a common receptor induced in tumor microvessels,cancer cells and cancer stem-like cells (CSCs) that is involved in tumor angiogenesis,invasiveness,and CSC anoikis resistance and survival,could underlie contemporaneous coordination of these events rather than assume stochasticity. Here we show that functional analysis of the dual endothelin1/VEGFsignal peptide receptor,DEspR,(formerly named Dear,Chr.4q31.2) supports the putative common receptor paradigm in pancreatic ductal adenocarcinoma (PDAC) and glioblastoma (GBM) selected for their invasiveness,CD133+CSCs,and polar angiogenic features. Unlike normal tissue,DEspR is detected in PDAC and GBM microvessels,tumor cells,and CSCs isolated from PDAC-Panc1 and GBM-U87 cells. DEspR-inhibition decreased angiogenesis,invasiveness,CSC-survival and anoikis resistance in vitro,and decreased Panc1-CSC and U87-CSC xenograft tumor growth,vasculo-angiogenesis and invasiveness in nude(nu/nu) rats,suggesting that DEspR activation would coordinate these tumor progression events. As an accessible,cell-surface 'common receptor coordinator',DEspR-inhibition defines a novel targeted-therapy paradigm for pancreatic cancer and glioblastoma. View Publication

产品号#：

05620

产品名：

MammoCult™ 人源培养基套装

Park Y et al. (MAR 2014) Journal of Biotechnology 174 1 39--48

Hepatic differentiation of human embryonic stem cells on microcarriers

Translation of stem cell research to industrial and clinical settings mostly requires large quantities of cells,especially those involving large organs such as the liver. A scalable reactor system is desirable to ensure a reliable supply of sufficient quantities of differentiated cells. To increase the culture efficiency in bioreactor system,high surface to volume ratio needs to be achieved. We employed a microcarrier culture system for the expansion of undifferentiated human embryonic stem cells (hESCs) as well as for directed differentiation of these cells to hepatocyte-like cells. Cells in single cell suspension were attached to the bead surface in even distribution and were expanded to 1??106cells/ml within 2 days of hESC culture with maintenance of the level of pluripotency markers. Directed differentiation into hepatocyte-like cells on microcarriers,both in static culture and stirred bioreactors,induced similar levels of hepatocyte-like cell differentiation as observed with cells cultured in conventional tissue culture plates. The cells expressed both immature and mature hepatocyte-lineage genes and proteins such as asialoglycoprotein receptor-1 (ASGPR-1) and albumin. Differentiated cells exhibited functional characteristics such as secretion of albumin and urea,and CYP3A4 activity could be detected. Microcarriers thus offer the potential for large-scale expansion and differentiation of hESCs induced hepatocyte-like cells in a more controllable bioreactor environment. ?? 2014. View Publication

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mTeSR™1

Kasper M et al. (FEB 1987) European journal of cancer & clinical oncology 23 2 137--47

Histological evaluation of three new monoclonal anti-cytokeratin antibodies. 1. Normal tissues.

Three new monoclonal anti-cytokeratin antibodies (mabs) potentially useful in cancer research and clinical diagnosis have been evaluated in immuno-histochemistry on cryostat sections of a broad variety of normal human tissues. A45-B/B3 reacts with all cells containing cytokeratins (epithelia and mesothelia). This mab positively identifies epithelial cells of any kind,and it may serve in differentiating carcinomas from tumours of mesenchymal origin. A53-B/A2 recognizes an individual cytokeratin,No. 19,and stains preferably mesothelia,urothelium,and bile duct epithelium. This antibody is suited to discriminate between different epithelial lineages. A51-B/H4 reacts with a subgroup of cytokeratins (probably including Nos. 14,8 and/or 18). It is positive with most epithelia but negative with keratanized stratified epithelium. This antibody shows an interesting,but up to now unexplained,cross-reactivity with nuclei of certain nonepithelial cells. All three mabs also react with epithelial cells from at least three animal species. View Publication

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Ikebe C and Suzuki K ( 2014) BioMed research international 2014 951512

Mesenchymal stem cells for regenerative therapy: optimization of cell preparation protocols.

Administration of bone marrow-derived mesenchymal stem cells (MSCs) is an innovative approach for the treatment of a range of diseases that are not curable by current therapies including heart failure. A number of clinical trials have been completed and many others are ongoing; more than 2,000 patients worldwide have been administered with culture-expanded allogeneic or autologous MSCs for the treatment of various diseases,showing feasibility and safety (and some efficacy) of this approach. However,protocols for isolation and expansion of donor MSCs vary widely between these trials,which could affect the efficacy of the therapy. It is therefore important to develop international standards of MSC production,which should be evidence-based,regulatory authority-compliant,of good medical practice grade,cost-effective,and clinically practical,so that this innovative approach becomes an established widely adopted treatment. This review article summarizes protocols to isolate and expand bone marrow-derived MSCs in 47 recent clinical trials of MSC-based therapy,which were published after 2007 onwards and provided sufficient methodological information. Identified issues and possible solutions associated with the MSC production methods,including materials and protocols for isolation and expansion,are discussed with reference to relevant experimental evidence with aim of future clinical success of MSC-based therapy. View Publication

产品号#：

07930

07931

07940

07955

07956

07959

07954

100-1061

07952

产品名：

CryoStor® CS10

Behar RZ et al. (MAR 2014) Toxicology in Vitro 28 2 198--208

Identification of toxicants in cinnamon-flavored electronic cigarette refill fluids

In a prior study on electronic cigarette (EC) refill fluids,Cinnamon Ceylon was the most cytotoxic of 36 products tested. The purpose of the current study was to determine if high cytotoxicity is a general feature of cinnamon-flavored EC refill fluids and to identify the toxicant(s) in Cinnamon Ceylon. Eight cinnamon-flavored refill fluids,which were screened using the MTT assay,varied in their cytotoxicity with most being cytotoxic. Human embryonic stem cells were generally more sensitive than human adult pulmonary fibroblasts. Most products were highly volatile and produced vapors that impaired survival of cells in adjacent wells. Cinnamaldehyde (CAD),2-methoxycinnamaldehyde (2MOCA),dipropylene glycol,and vanillin were identified in the cinnamon-flavored refill fluids using gas chromatography-mass spectrometry and high-pressure liquid chromatography (HPLC). When authentic standards of each chemical were tested using the MTT assay,only CAD and 2MOCA were highly cytotoxic. The amount of each chemical in the refill fluids was quantified using HPLC,and cytotoxicity correlated with the amount of CAD/product. Duplicate bottles of the same product were similar,but varied in their concentrations of 2MOCA. These data show that the cinnamon flavorings in refill fluids are linked to cytotoxicity,which could adversely affect EC users. ?? 2013 Elsevier Ltd. View Publication

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mTeSR™1

Chronopoulou E et al. ( 2014) 1131 47--70

Hybridoma technology for the generation of rodent mAbs via classical fusion

Monoclonal antibodies (mAbs) have proven to be instrumental in the advancement of research,diagnostic,industrial vaccine,and therapeutic applications. The use of mAbs in laboratory protocols has been growing in an exponential fashion for the last four decades. Described herein are methods for the development of highly specific mAbs through traditional hybridoma fusion. For ultimate success,a series of simultaneously initiated protocols are to be undertaken with careful attention to cell health of both the myeloma fusion partner and immune splenocytes. Coordination and attention to detail will enable a researcher with basic tissue culture skills to generate mAbs from immunized rodents to a variety of antigens (including proteins,carbohydrates,DNA,and haptens) (see Note 1). Furthermore,in vivo and in vitro methods used for antigen sensitization of splenocytes prior to somatic fusion are described herein. View Publication

产品号#：

03800

03801

03802

03803

03804

03805

03806

产品名：

ClonaCell™-HY杂交瘤试剂盒

ClonaCell™-HY培养基A

ClonaCell™-HY 培养基 B

ClonaCell™-HY 培养基 C

ClonaCell™-HY 培养基 D

ClonaCell™-HY 培养基 E

ClonaCell™-HY PEG

Bernet JD et al. (MAR 2014) Nature medicine 20 3 265--71

p38 MAPK signaling underlies a cell-autonomous loss of stem cell self-renewal in skeletal muscle of aged mice.

Skeletal muscle aging results in a gradual loss of skeletal muscle mass,skeletal muscle function and regenerative capacity,which can lead to sarcopenia and increased mortality. Although the mechanisms underlying sarcopenia remain unclear,the skeletal muscle stem cell,or satellite cell,is required for muscle regeneration. Therefore,identification of signaling pathways affecting satellite cell function during aging may provide insights into therapeutic targets for combating sarcopenia. Here,we show that a cell-autonomous loss in self-renewal occurs via alterations in fibroblast growth factor receptor-1,p38α and p38β mitogen-activated protein kinase signaling in satellite cells from aged mice. We further demonstrate that pharmacological manipulation of these pathways can ameliorate age-associated self-renewal defects. Thus,our data highlight an age-associated deregulation of a satellite cell homeostatic network and reveal potential therapeutic opportunities for the treatment of progressive muscle wasting. View Publication

产品号#：

72682

72684

产品名：

BIRB - 796

Sousa-Ferreira L et al. ( 2014) PloS one 9 3 e88917

Fluoxetine induces proliferation and inhibits differentiation of hypothalamic neuroprogenitor cells in vitro.

A significant number of children undergo maternal exposure to antidepressants and they often present low birth weight. Therefore,it is important to understand how selective serotonin reuptake inhibitors (SSRIs) affect the development of the hypothalamus,the key center for metabolism regulation. In this study we investigated the proliferative actions of fluoxetine in fetal hypothalamic neuroprogenitor cells and demonstrate that fluoxetine induces the proliferation of these cells,as shown by increased neurospheres size and number of proliferative cells (Ki-67+ cells). Moreover,fluoxetine inhibits the differentiation of hypothalamic neuroprogenitor cells,as demonstrated by decreased number of mature neurons (Neu-N+ cells) and increased number of undifferentiated cells (SOX-2+ cells). Additionally,fluoxetine-induced proliferation and maintenance of hypothalamic neuroprogenitor cells leads to changes in the mRNA levels of appetite regulator neuropeptides,including Neuropeptide Y (NPY) and Cocaine-and-Amphetamine-Regulated-Transcript (CART). This study provides the first evidence that SSRIs affect the development of hypothalamic neuroprogenitor cells in vitro with consequent alterations on appetite neuropeptides. View Publication

产品号#：

73142

73144

产品名：

Liu B et al. (MAR 2014) PLoS ONE 9 3 e90615

Nanog1 in NTERA-2 and recombinant NanogP8 from somatic cancer cells adopt multiple protein conformations and migrate at multiple M.W species

Human Nanog1 is a 305-amino acid (aa) homeodomain-containing transcription factor critical for the pluripotency of embryonic stem (ES) and embryonal carcinoma (EC) cells. Somatic cancer cells predominantly express a retrogene homolog of Nanog1 called NanogP8,which is ˜99% similar to Nanog at the aa level. Although the predicted M.W of Nanog1/NanogP8 is ∼35 kD,both have been reported to migrate,on Western blotting (WB),at apparent molecular masses of 29-80 kD. Whether all these reported protein bands represent authentic Nanog proteins is unclear. Furthermore,detailed biochemical studies on Nanog1/NanogpP8 have been lacking. By combining WB using 8 anti-Nanog1 antibodies,immunoprecipitation,mass spectrometry,and studies using recombinant proteins,here we provide direct evidence that the Nanog1 protein in NTERA-2 EC cells exists as multiple M.W species from ˜22 kD to 100 kD with a major 42 kD band detectable on WB. We then demonstrate that recombinant NanogP8 (rNanogP8) proteins made in bacteria using cDNAs from multiple cancer cells also migrate,on denaturing SDS-PAGE,at ˜28 kD to 180 kD. Interestingly,different anti-Nanog1 antibodies exhibit differential reactivity towards rNanogP8 proteins,which can spontaneously form high M.W protein species. Finally,we show that most long-term cultured cancer cell lines seem to express very low levels of or different endogenous NanogP8 protein that cannot be readily detected by immunoprecipitation. Altogether,the current study reveals unique biochemical properties of Nanog1 in EC cells and NanogP8 in somatic cancer cells. View Publication

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mTeSR™1

Sareen D et al. (AUG 2014) Journal of Comparative Neurology 522 12 2707--2728

Human induced pluripotent stem cells are a novel source of neural progenitor cells (iNPCs) that migrate and integrate in the rodent spinal cord

Transplantation of human neural progenitor cells (NPCs) into the brain or spinal cord to replace lost cells,modulate the injury environment,or create a permissive milieu to protect and regenerate host neurons is a promising therapeutic strategy for neurological diseases. Deriving NPCs from human fetal tissue is feasible,although problematic issues include limited sources and ethical concerns. Here we describe a new and abundant source of NPCs derived from human induced pluripotent stem cells (iPSCs). A novel chopping technique was used to transform adherent iPSCs into free-floating spheres that were easy to maintain and were expandable (EZ spheres) (Ebert et al. [2013] Stem Cell Res 10:417–427). These EZ spheres could be differentiated towards NPC spheres with a spinal cord phenotype using a combination of all-trans retinoic acid (RA) and epidermal growth factor (EGF) and fibroblast growth factor-2 (FGF-2) mitogens. Suspension cultures of NPCs derived from human iPSCs or fetal tissue have similar characteristics,although they were not similar when grown as adherent cells. In addition,iPSC-derived NPCs (iNPCs) survived grafting into the spinal cord of athymic nude rats with no signs of overgrowth and with a very similar profile to human fetal-derived NPCs (fNPCs). These results suggest that human iNPCs behave like fNPCs and could thus be a valuable alternative for cellular regenerative therapies of neurological diseases. J. Comp. Neurol. 522:2707–2728,2014. textcopyright 2014 Wiley Periodicals,Inc. View Publication

产品号#：

05850

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mTeSR™1

技术资料

OCT4 as a target of miR-34a stimulates p63 but inhibits p53 to promote human cell transformation

Assessment of the purity of isolated cell populations for lineage-specific chimerism monitoring post haematopoietic stem cell transplantation.

DEspR roles in tumor vasculo-angiogenesis, invasiveness, CSC-survival and anoikis resistance: a 'common receptor coordinator' paradigm.

Hepatic differentiation of human embryonic stem cells on microcarriers

Histological evaluation of three new monoclonal anti-cytokeratin antibodies. 1. Normal tissues.

Mesenchymal stem cells for regenerative therapy: optimization of cell preparation protocols.

Identification of toxicants in cinnamon-flavored electronic cigarette refill fluids

Hybridoma technology for the generation of rodent mAbs via classical fusion

p38 MAPK signaling underlies a cell-autonomous loss of stem cell self-renewal in skeletal muscle of aged mice.

Fluoxetine induces proliferation and inhibits differentiation of hypothalamic neuroprogenitor cells in vitro.

Nanog1 in NTERA-2 and recombinant NanogP8 from somatic cancer cells adopt multiple protein conformations and migrate at multiple M.W species

Human induced pluripotent stem cells are a novel source of neural progenitor cells (iNPCs) that migrate and integrate in the rodent spinal cord

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