Isolation of a Monoclonal Human scFv Against Cytomegalovirus pp71 Antigen Using Yeast Display
Background: Human cytomegalovirus (CMV) is a major pathogen that poses significant risks to immunocompromised individuals and neonates. The tegument protein pp71,encoded by the UL82 gene,plays a pivotal role in initiating viral lytic replication and evading host immune responses. Despite its clinical relevance,standardized monoclonal antibodies (mAbs) for pp71 remain limited,prompting the need to expand the available repertoire of antibodies targeting this critical protein. Methods: In this study,we constructed a diverse human single-chain variable fragment (scFv) library using RNA derived from the B cells of four healthy donors. The library was expressed in Saccharomyces cerevisiae,and iterative rounds of magnetic-activated cell sorting (MACS) were performed against recombinant pp71. Clonal enrichment was monitored using flow cytometry. Results: Among the isolated clones,one designated ID2 exhibited high sensitivity and specificity for pp71,as demonstrated by flow cytometry,immunofluorescence,an enzyme-linked immunosorbent assay (ELISA),and biolayer interferometry (BLI). Conclusions: Collectively,these findings establish a novel pp71-specific mAb and underscore the utility of yeast surface display combined with MACS for expanding the antibody toolkit available for CMV research and diagnostics.
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产品类型：
产品号#：
100-0645
产品名：
ImmunoCult™ 人B细胞扩增试剂盒
Leberbauer C et al. (JAN 2005)
Blood 105 1 85--94
Different steroids co-regulate long-term expansion versus terminal differentiation in primary human erythroid progenitors.
Outgrowth,long-term self-renewal,and terminal maturation of human erythroid progenitors derived from umbilical cord blood in serum-free medium can be modulated by steroid hormones. Homogeneous erythroid cultures,as characterized by flow cytometry and dependence on a specific mixture of physiologic proliferation factors,were obtained within 8 days from a starting population of mature and immature mononuclear cells. Due to previous results in mouse and chicken erythroblasts,the proliferation-promoting effect of glucocorticoids was not unexpected. Surprisingly,however,androgen had a positive effect on the sustained expansion of human female but not male erythroid progenitors. Under optimal conditions,sustained proliferation of erythroid progenitors resulted in a more than 10(9)-fold expansion within 60 days. Terminal erythroid maturation was significantly improved by adding human serum and thyroid hormone (3,5,3'-triiodothyronine [T3]) to the differentiation medium. This resulted in highly synchronous differentiation of the cells toward enucleated erythrocytes within 6 days,accompanied by massive size decrease and hemoglobin accumulation to levels comparable to those in peripheral blood erythrocytes. Thus,obviously,different ligand-activated nuclear hormone receptors massively influence the decision between self-renewal and terminal maturation in the human erythroid compartment.
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Production of de novo cardiomyocytes: human pluripotent stem cell differentiation and direct reprogramming.
Cardiovascular disease is a leading cause of death worldwide. The limited capability of heart tissue to regenerate has prompted methodological developments for creating de novo cardiomyocytes,both in vitro and in vivo. Beyond uses in cell replacement therapy,patient-specific cardiomyocytes may find applications in drug testing,drug discovery,and disease modeling. Recently,approaches for generating cardiomyocytes have expanded to encompass three major sources of starting cells: human pluripotent stem cells (hPSCs),adult heart-derived cardiac progenitor cells (CPCs),and reprogrammed fibroblasts. We discuss state-of-the-art methods for generating de novo cardiomyocytes from hPSCs and reprogrammed fibroblasts,highlighting potential applications and future challenges.
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产品类型：
产品号#：
05850
05857
05870
05875
85850
85857
85870
85875
产品名：
mTeSR™1
mTeSR™1
Liu M-L et al. (JAN 2013)
Nature communications 4 2183
Small molecules enable neurogenin 2 to efficiently convert human fibroblasts into cholinergic neurons.
Cell fate can be reprogrammed by modifying intrinsic and extrinsic cues. Here we show that two small molecules (forskolin and dorsomorphin) enable the transcription factor Neurogenin 2 (NGN2) to convert human fetal lung fibroblasts into cholinergic neurons with high purity (textgreater90%) and efficiency (up to 99% of NGN2-expressing cells). The conversion is direct without passing through a proliferative progenitor state. These human induced cholinergic neurons (hiCN) show mature electrophysiological properties and exhibit motor neuron-like features,including morphology,gene expression and the formation of functional neuromuscular junctions. Inclusion of an additional transcription factor,SOX11,also efficiently converts postnatal and adult skin fibroblasts from healthy and diseased human patients to cholinergic neurons. Taken together,this study identifies a simple and highly efficient strategy for reprogramming human fibroblasts to subtype-specific neurons. These findings offer a unique venue for investigating the molecular mechanisms underlying cellular plasticity and human neurodegenerative diseases.
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产品类型：
产品号#：
72112
72114
100-0249
产品名：
Forskolin
Forskolin
Forskolin
Mora-Bermú et al. (SEP 2016)
eLife 5
Differences and similarities between human and chimpanzee neural progenitors during cerebral cortex development.
Human neocortex expansion likely contributed to the remarkable cognitive abilities of humans. This expansion is thought to primarily reflect differences in proliferation versus differentiation of neural progenitors during cortical development. Here,we have searched for such differences by analysing cerebral organoids from human and chimpanzees using immunohistochemistry,live imaging,and single-cell transcriptomics. We find that the cytoarchitecture,cell type composition,and neurogenic gene expression programs of humans and chimpanzees are remarkably similar. Notably,however,live imaging of apical progenitor mitosis uncovered a lengthening of prometaphase-metaphase in humans compared to chimpanzees that is specific to proliferating progenitors and not observed in non-neural cells. Consistent with this,the small set of genes more highly expressed in human apical progenitors points to increased proliferative capacity,and the proportion of neurogenic basal progenitors is lower in humans. These subtle differences in cortical progenitors between humans and chimpanzees may have consequences for human neocortex evolution.
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产品类型：
产品号#：
05850
05857
05870
05875
85850
85857
85870
85875
产品名：
mTeSR™1
mTeSR™1
Pabst C et al. (APR 2014)
Nature methods 11 4 436--42
Identification of small molecules that support human leukemia stem cell activity ex vivo.
Leukemic stem cells (LSCs) are considered a major cause of relapse in acute myeloid leukemia (AML). Defining pathways that control LSC self-renewal is crucial for a better understanding of underlying mechanisms and for the development of targeted therapies. However,currently available culture conditions do not prevent spontaneous differentiation of LSCs,which greatly limits the feasibility of cell-based assays. To overcome these constraints we conducted a high-throughput chemical screen and identified small molecules that inhibit differentiation and support LSC activity in vitro. Similar to reports with cord blood stem cells,several of these compounds suppressed the aryl-hydrocarbon receptor (AhR) pathway,which we show to be inactive in vivo and rapidly activated ex vivo in AML cells. We also identified a compound,UM729,that collaborates with AhR suppressors in preventing AML cell differentiation. Together,these findings provide newly defined culture conditions for improved ex vivo culture of primary human AML cells.
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产品类型：
产品号#：
72332
72334
产品名：
UM729
Freedman BS et al. (OCT 2015)
Nature communications 6 May 8715
Modelling kidney disease with CRISPR-mutant kidney organoids derived from human pluripotent epiblast spheroids.
Human-pluripotent-stem-cell-derived kidney cells (hPSC-KCs) have important potential for disease modelling and regeneration. Whether the hPSC-KCs can reconstitute tissue-specific phenotypes is currently unknown. Here we show that hPSC-KCs self-organize into kidney organoids that functionally recapitulate tissue-specific epithelial physiology,including disease phenotypes after genome editing. In three-dimensional cultures,epiblast-stage hPSCs form spheroids surrounding hollow,amniotic-like cavities. GSK3β inhibition differentiates spheroids into segmented,nephron-like kidney organoids containing cell populations with characteristics of proximal tubules,podocytes and endothelium. Tubules accumulate dextran and methotrexate transport cargoes,and express kidney injury molecule-1 after nephrotoxic chemical injury. CRISPR/Cas9 knockout of podocalyxin causes junctional organization defects in podocyte-like cells. Knockout of the polycystic kidney disease genes PKD1 or PKD2 induces cyst formation from kidney tubules. All of these functional phenotypes are distinct from effects in epiblast spheroids,indicating that they are tissue specific. Our findings establish a reproducible,versatile three-dimensional framework for human epithelial disease modelling and regenerative medicine applications.
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产品类型：
产品号#：
05850
05857
05870
05875
07920
85850
85857
85870
85875
07922
产品名：
ACCUTASE™
mTeSR™1
mTeSR™1
ACCUTASE™
Y.-W. Liu et al. (AUG 2018)
Nature biotechnology 36 7 597--605
Human embryonic stem cell-derived cardiomyocytes restore function in infarcted hearts of non-human primates.
Pluripotent stem cell-derived cardiomyocyte grafts can remuscularize substantial amounts of infarcted myocardium and beat in synchrony with the heart,but in some settings cause ventricular arrhythmias. It is unknown whether human cardiomyocytes can restore cardiac function in a physiologically relevant large animal model. Here we show that transplantation of ∼750 million cryopreserved human embryonic stem cell-derived cardiomyocytes (hESC-CMs) enhances cardiac function in macaque monkeys with large myocardial infarctions. One month after hESC-CM transplantation,global left ventricular ejection fraction improved 10.6 ± 0.9{\%} vs. 2.5 ± 0.8{\%} in controls,and by 3 months there was an additional 12.4{\%} improvement in treated vs. a 3.5{\%} decline in controls. Grafts averaged 11.6{\%} of infarct size,formed electromechanical junctions with the host heart,and by 3 months contained ∼99{\%} ventricular myocytes. A subset of animals experienced graft-associated ventricular arrhythmias,shown by electrical mapping to originate from a point-source acting as an ectopic pacemaker. Our data demonstrate that remuscularization of the infarcted macaque heart with human myocardium provides durable improvement in left ventricular function.
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产品类型：
产品号#：
07930
07931
07940
07955
07959
07952
100-1061
产品名：
CryoStor® CS10
CryoStor® CS10
CryoStor® CS10
CryoStor® CS10
CryoStor® CS10
CryoStor® CS10
CryoStor® CS10
I. M. Saldana-Guerrero et al. (May 2024)
Nature Communications 15
A human neural crest model reveals the developmental impact of neuroblastoma-associated chromosomal aberrations
Early childhood tumours arise from transformed embryonic cells,which often carry large copy number alterations (CNA). However,it remains unclear how CNAs contribute to embryonic tumourigenesis due to a lack of suitable models. Here we employ female human embryonic stem cell (hESC) differentiation and single-cell transcriptome and epigenome analysis to assess the effects of chromosome 17q/1q gains,which are prevalent in the embryonal tumour neuroblastoma (NB). We show that CNAs impair the specification of trunk neural crest (NC) cells and their sympathoadrenal derivatives,the putative cells-of-origin of NB. This effect is exacerbated upon overexpression of MYCN,whose amplification co-occurs with CNAs in NB. Moreover,CNAs potentiate the pro-tumourigenic effects of MYCN and mutant NC cells resemble NB cells in tumours. These changes correlate with a stepwise aberration of developmental transcription factor networks. Together,our results sketch a mechanistic framework for the CNA-driven initiation of embryonal tumours. Subject terms: Paediatric cancer,Stem cells,Disease model,Cancer genomics,Embryonal neoplasms
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产品类型：
产品号#：
05790
产品名：
BrainPhys™神经元培养基
S-Y. Kim et al. (Feb 2026)
Materials Today Bio 37 8
CRISPR-engineered human lung organoids with a biomolecular condensate reporter enable mechanistic toxicity monitoring
Understanding how chemical stress perturbs human lung physiology requires models that capture dynamic molecular responses in real time. Here,we established a CRISPR/Cas9-engineered human induced pluripotent stem cell (hiPSC)-derived lung organoid expressing endogenous G3BP1–mCherry,enabling live,non-destructive visualization of stress granule (SG) formation under toxicant exposure. The organoids recapitulated airway and alveolar epithelial diversity and displayed lamellar body-like ultrastructures,indicating advanced maturation. Time-lapse imaging revealed rapid and reversible SG dynamics across chemically distinct stressors,while cytotoxicity assays showed that these organoids are significantly more sensitive than conventional 2D or cancer-derived lung models. Importantly,SG dynamics were linked to exposure duration–dependent changes in epithelial barrier integrity,indicating that SG formation precedes overt epithelial injury and serves as an early indicator of toxicant-induced cellular stress. Integration with high-content screening enabled quantitative,image-based analysis of cellular stress phenotypes,greatly enhancing throughput and mechanistic insight,thereby provided next-generation New Approach Methodologies for lung toxicity assessment. Together,this hiPSC-derived lung organoid SG reporter platform links early molecular stress adaptation to tissue-level responses,offering a predictive and mechanistically informative framework for human-relevant lung toxicity evaluation.
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产品类型：
产品号#：
05110
05946
34411
34415
34421
34425
34450
34460
85850
85857
产品名：
STEMdiff™定型内胚层检测试剂盒
TeSR™-E6
AggreWell™ 400 24孔板，1个
AggreWell™400 24孔板，5个
AggreWell™ 400 6孔板，1个
AggreWell™ 400 6孔板，5个
AggreWell™400 24孔板启动套装
AggreWell™ 400 6孔板启动套装
mTeSR™1
mTeSR™1
(Feb 2024)
Cell Death & Disease 15 2
CUL4B mutations impair human cortical neurogenesis through PP2A-dependent inhibition of AKT and ERK
Mutation in CUL4B gene is one of the most common causes for X-linked intellectual disability (XLID). CUL4B is the scaffold protein in CUL4B-RING ubiquitin ligase (CRL4B) complex. While the roles of CUL4B in cancer progression and some developmental processes like adipogenesis,osteogenesis,and spermatogenesis have been studied,the mechanisms underlying the neurological disorders in patients with CUL4B mutations are poorly understood. Here,using 2D neuronal culture and cerebral organoids generated from the patient-derived induced pluripotent stem cells and their isogenic controls,we demonstrate that CUL4B is required to prevent premature cell cycle exit and precocious neuronal differentiation of neural progenitor cells. Moreover,loss-of-function mutations of CUL4B lead to increased synapse formation and enhanced neuronal excitability. Mechanistically,CRL4B complex represses transcription of PPP2R2B and PPP2R2C genes,which encode two isoforms of the regulatory subunit of protein phosphatase 2 A (PP2A) complex,through catalyzing monoubiquitination of H2AK119 in their promoter regions. CUL4B mutations result in upregulated PP2A activity,which causes inhibition of AKT and ERK,leading to premature cell cycle exit. Activation of AKT and ERK or inhibition of PP2A activity in CUL4B mutant organoids rescues the neurogenesis defect. Our work unveils an essential role of CUL4B in human cortical development.
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产品类型：
产品号#：
85850
85857
产品名：
mTeSR™1
mTeSR™1
(Apr 2024)
Nature Communications 15
Complex activity and short-term plasticity of human cerebral organoids reciprocally connected with axons
An inter-regional cortical tract is one of the most fundamental architectural motifs that integrates neural circuits to orchestrate and generate complex functions of the human brain. To understand the mechanistic significance of inter-regional projections on development of neural circuits,we investigated an in vitro neural tissue model for inter-regional connections,in which two cerebral organoids are connected with a bundle of reciprocally extended axons. The connected organoids produced more complex and intense oscillatory activity than conventional or directly fused cerebral organoids,suggesting the inter-organoid axonal connections enhance and support the complex network activity. In addition,optogenetic stimulation of the inter-organoid axon bundles could entrain the activity of the organoids and induce robust short-term plasticity of the macroscopic circuit. These results demonstrated that the projection axons could serve as a structural hub that boosts functionality of the organoid-circuits. This model could contribute to further investigation on development and functions of macroscopic neuronal circuits in vitro. Connecting cerebral organoids with an axon bundle models inter-regional projections and enhances neural activity. Optogenetic stimulation induces short-term plasticity,offering insights into macroscopic circuit development and functionality.
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EasySep™小鼠TIL（CD45）正选试剂盒
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