Abstract:
:Functional imaging of the intracellular calcium concentration [Ca2+]i using fluorescent indicators is a powerful and frequently applied method for assessing various biological questions in vitro, including ion channel function and intracellular signaling in homeostasis and disease. In functional [Ca2+]i imaging experiments, the fluorescence intensity of single cells is typically recorded during application of a chemical stimulus, i.e. by exchange of modified extracellular media, exposure to drugs and/or ligands. The concomitant mechanical perturbation caused by the perfusion of different solution during experimentation severely hinders calcium imaging in non-adherent cells, including peripheral immune cells, as cells in suspension are dislocated by turbulent flow during chemical stimulation. The quantitative analysis, involving time-courses of intracellular fluorescence signal changes, necessitates cells to remain at the same position throughout the experiment. To prevent dislocation of cells during solution exchange, and to enable imaging as well as analysis of Ca2+ responses in immune cells, a gelatin-based method for immobilization of non-adherent cells was developed. Gelatin has been a long-serving material for cell immobilization, e.g. in 3D bio-printing of cells and has thus, also been employed in the context of this study. To demonstrate the applicability of the established method for functional Ca2+ imaging in gelatin-immobilized suspension cells, a proof-of-concept study was conducted using human peripheral blood model cell lines (Jurkat/T-lymphocytes and THP-1/monocytes), Ca2+ indicators (Fluo-4 and Fura-2) and two different fluorescence microscopy rigs. The data presented that the established methodology is applicable for studying Ca2+ signaling by in vitro high-content functional imaging of [Ca2+]i in suspension cells, including but not restricted to human immune cells.
journal_name
Exp Cell Resjournal_title
Experimental cell researchauthors
Ritter P,Bye LJ,Finol-Urdaneta RK,Lesko C,Adams DJ,Friedrich O,Gilbert DFdoi
10.1016/j.yexcr.2020.112210subject
Has Abstractpub_date
2020-10-15 00:00:00pages
112210issue
2eissn
0014-4827issn
1090-2422pii
S0014-4827(20)30459-6journal_volume
395pub_type
杂志文章abstract::Keratinocyte growth factor (KGF/FGF7) and fibroblast growth factor 10 (FGF10/KGF2) regulate keratinocyte proliferation and differentiation by binding to the tyrosine kinase KGF receptor (KGFR). KGF induces keratinocyte motility and cytoskeletal rearrangement, whereas a direct role of FGF10 on keratinocyte migration is...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/j.yexcr.2007.03.013
更新日期:2007-05-15 00:00:00
abstract::Molecular studies of the physiological cell death process have focused attention on the role of effector caspases as critical common elements of the lethal mechanism. Diverse death signals act afferently via distinct signaling pathways to activate these resident proenzyme molecules post-translationally. Whether this m...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/s0014-4827(03)00289-1
更新日期:2003-10-01 00:00:00
abstract::The Cdc25 phosphatases play crucial roles in cell cycle progression by removing inhibitory phosphates from tyrosine and threonine residues of cyclin-dependent kinases. Cdc25A is an important regulator of the G1/S transition but functions also in the mitotic phase of the human cell cycle. In this paper, we investigate ...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/j.yexcr.2004.09.012
更新日期:2005-02-01 00:00:00
abstract::The pre-mRNA splicing factor 4 kinase PRP4K (PRPF4B), is an essential kinase that is a component of the U5 snRNP and functions in spliceosome assembly. We demonstrated that PRP4K is a novel biological marker for taxane response in ovarian cancer patients and reduced levels of PRP4K correlate with intrinsic and acquire...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/j.yexcr.2015.12.013
更新日期:2016-01-01 00:00:00
abstract::Tpr is a 267-kDa protein of unknown function recently identified as a constitutive component of the nuclear pore complex (NPC2)-attached intranuclear filaments. Secondary structure predictions suggest that the protein is divided into a large, coiled-coil forming aminoterminal domain and a shorter, highly acidic carbox...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1006/excr.1998.4246
更新日期:1998-11-25 00:00:00
abstract::Celiac disease (CD) is a multisystemic autoimmune inflammation of the intestinal tract induced by wheat gluten and related cereals in HLA-DQ2/8 positive individuals. An essential role in the pathogenesis of CD is played by a fraction of the peptic-tryptic digest of gluten, Frazer's Fraction (FF). Here, we investigate ...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/j.yexcr.2009.06.026
更新日期:2009-11-15 00:00:00
abstract::Mammalian enabled (MENA), usually known as a direct regulator of microfilament polymerization and bundling, promotes metastasis in various cancers. Here we focus on the role of MENA in hepatocellular carcinoma (HCC) metastasis and the relevant mechanism from the view of RhoA activity regulation. By HCC tissue microarr...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/j.yexcr.2014.05.010
更新日期:2014-09-10 00:00:00
abstract::Insulin-like growth factor-1 (IGF-1) and its receptor (IGF-1R) are required for cell proliferation in vitro, raising the possibility that an autocrine IGF-1-IGF-1R system may be present in vivo and become activated in the viable ventricular myocytes shortly after infarction. Therefore, following the in vivo documentat...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1006/excr.1994.1224
更新日期:1994-08-01 00:00:00
abstract::Spinal muscular atrophy (SMA), a recessive, neuromuscular disease, is caused by a mutation or deletion in the SMN1 gene. The SMN2 gene is present in the same region of chromosome 5 and is similar in DNA sequence to SMN1 except for a T at position +6 of exon 7 that is likely the predominant functional difference betwee...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/j.yexcr.2007.10.012
更新日期:2008-02-15 00:00:00
abstract::Many recent studies have revealed that axin is involved in numerous cellular functions beyond the negative regulation of β-catenin-dependent Wnt signaling. Previously, an association of ectopic axin with mitochondria was observed. In an effort to investigate the relationship between axin and mitochondria, we found tha...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/j.yexcr.2015.12.003
更新日期:2016-01-01 00:00:00
abstract::The Ras-like family of small GTPases includes, among others, Ras, Rap1, R-ras, and Ral. The family is characterized by similarities in the effector domain. While the function of Ras is, at least in part, elucidated, little is known about other members of the family. Currently, much attention is focused on the small GT...
journal_title:Experimental cell research
pub_type: 杂志文章,评审
doi:10.1006/excr.1999.4695
更新日期:1999-11-25 00:00:00
abstract::Bax∆2 is a functional pro-apoptotic Bax isoform having alterations in its N-terminus, but sharing the rest of its sequence with Baxα. Bax∆2 is unable to target mitochondria due to the loss of helix α1. Instead, it forms cytosolic aggregates and activates caspase 8. However, the functional domain(s) responsible for Bax...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/j.yexcr.2017.08.016
更新日期:2017-10-15 00:00:00
abstract:BACKGROUND:Metastatic and/or recurrent breast carcinomas are leading causes of cancer-related death worldwide. Breast cancer stem cells (BCSCs) have been implicated in cancer metastases and progression, thus, the need for the discovery and development of effective BCSCs-specific therapies against metastatic and triple ...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/j.yexcr.2018.07.008
更新日期:2018-09-15 00:00:00
abstract::Abdominal aortic aneurysms (AAA) are characterized by matrix remodeling, elastin degradation, absence of nitric oxide (NO) signaling, and inflammation, influencing smooth muscle cell (SMC) phenotype and gene expression. Little is known about the biomolecular release and intrinsic biomechanics of human AAA-SMCs. NO del...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/j.yexcr.2019.111589
更新日期:2019-11-01 00:00:00
abstract::The mammalian Hippo pathway comprises mammalian Ste20-like kinases (MST1/2) and large tumor suppressor kinases (LATS1/2). LATS1/2, which are activated by MST1/2, phosphorylate a transcriptional co-activator, yes-associated protein (YAP), and induce the recruitment of YAP by 14-3-3 to cytoplasm, so that the TEAD-depend...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/j.yexcr.2013.01.020
更新日期:2013-04-15 00:00:00
abstract::We established a differentiation method for homogeneous α7 integrin-positive human skeletal muscle stem cell (α7(+)hSMSC)-derived osteoblast-like cells with bone morphogenetic protein (BMP)-2. To explore the early signaling cascade for osteoblastic differentiation, we examined the upregulation of autophagy-related gen...
journal_title:Experimental cell research
pub_type: 杂志文章,收录出版
doi:10.1016/j.yexcr.2016.07.002
更新日期:2016-09-10 00:00:00
abstract::Specific cholecystokinin (CCK) receptor and gastrin receptor antagonists were used to assess what role, if any, these receptors have in autocrine cell growth. Although the cholecystokinin receptor antagonist, L-364,718, inhibited cell proliferation in a broad spectrum of cell lines, the gastrin antagonist, L-365,260, ...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/0014-4827(91)90174-s
更新日期:1991-01-01 00:00:00
abstract::The intranuclear localization of SV40 T-antigen (T-Ag) and the cellular protein p53 was studied in SV40 abortively infected baby mouse kidney cells using two complementary methods of ultrastructural immunocytochemistry in combination with preferential staining of nuclear RNP components and electron microscope autoradi...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/0014-4827(88)90026-2
更新日期:1988-07-01 00:00:00
abstract::Mitochondrial DNA (mtDNA) is a multicopy, maternally inherited, genome. Individuals frequently carry a mixture of genetically distinct mtDNA molecules whose proportions may vary between sexual generations or among tissues from the same individual. Analyses of the genetic composition of mitochondria have previously rel...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1006/excr.2000.4949
更新日期:2000-08-25 00:00:00
abstract::Transcription factor Foxa1 plays a critical role during neural differentiation and is induced immediately after retinoic acid (RA)-initiated differentiation of pluripotent P19 embryonal carcinoma cells, correlated with the downregulated expression of pluripotency-related genes such as Nanog. To study whether Foxa1 par...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/j.yexcr.2014.04.020
更新日期:2014-08-15 00:00:00
abstract::Tendinopathy is a common and disabling condition that is difficult to treat. The pathomolecular events behind tendinopathy remain uncertain. Micro-RNAs (miRNAs, miRs) are short non-coding RNAs that regulate gene expression and may play a role in tendinopathy development. Tenocytes were obtained from human patellar ten...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/j.yexcr.2019.03.008
更新日期:2019-05-15 00:00:00
abstract::The activation of cdc2/cyclin B is the trigger for entry into mitosis. The mechanism of cdc2/cyclin B activation is complex, but the final step is the dephosphorylation of the Thr14 and Tyr15 residues on the cdc2 subunit, catalyzed by a member of the Cdc25 family of phosphatases. Cdc2/cyclin B1 accumulates at the cent...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1006/excr.2000.4872
更新日期:2000-05-25 00:00:00
abstract::The laminins consist of at least 11 polypeptides (5 alpha-chains, 3 beta-chains, and 3 gamma-chains) specific to basement membranes. Here we investigate the biological activity associated with the G domain of the newly identified laminin alpha5-chain using 113 overlapping synthetic peptides (positions 2679-3635). Usin...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1006/excr.2002.5540
更新日期:2002-07-01 00:00:00
abstract::Elastin, a major constituent of the media, may play an important role in the attachment, migration, and phenotypic properties of vascular smooth muscle cells (SMC). We examined binding proteins and binding sites on elastin in rabbit arterial SMC in primary culture. Freshly isolated cells on alpha-elastin underwent phe...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1006/excr.1995.1164
更新日期:1995-05-01 00:00:00
abstract::Mixed lineage leukemia (MLL) arises from several KMT2A-gene chromosomal translocations. Shb gene deficiency has been found to exhibit pleiotropic effects in different models of leukemia, and consequently, this study aimed to investigate MLL-AF9-induced leukemia in Shb deficiency. Bone marrow cells from wild type and S...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/j.yexcr.2020.112368
更新日期:2020-12-15 00:00:00
abstract::The ChlR1 DNA helicase, encoded by DDX11 gene, which is responsible for Warsaw breakage syndrome (WABS), has a role in sister-chromatid cohesion. In this study, we show that human ChlR1 deficient cells exhibit abnormal heterochromatin organization. While constitutive heterochromatin is discretely localized at perinucl...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/j.yexcr.2011.08.006
更新日期:2011-10-15 00:00:00
abstract::hnRNP I, also referred to as polypyrimidine tract binding protein, is one of the proteins associated with nascent pre-mRNA in the heterogeneous nuclear ribonucleoprotein (hnRNP) complexes. As for all karyophilic proteins, the nuclear import of hnRNP proteins requires specific sequence determinants that in many instanc...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1006/excr.1997.3677
更新日期:1997-08-25 00:00:00
abstract::Both hypoxia and overexpression of the urokinase plasminogen activator receptor (uPAR) are associated with a poor clinical outcome in human cancers. Hypoxia has been shown to induce uPAR expression in breast cancer cells and to increase their invasion through Matrigel, a phenomenon which can be blocked using an anti-u...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1006/excr.1999.4804
更新日期:2000-03-15 00:00:00
abstract::Polycystin-2 (PC2, TRPP2) is a nonselective cation channel whose dysfunction is associated with the onset of autosomal dominant polycystic kidney disease (ADPKD). PC2 contributes to Ca2+ transport and cell signaling in renal epithelia and other tissues. Little is known however, as to the external Ca2+ regulation of PC...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/j.yexcr.2016.11.004
更新日期:2017-01-01 00:00:00
abstract::Rat myotubes in tissue culture form broad areas of close contact with the substrate. These areas often display two distinct, interdigitating sets of membrane domains. One, the "contact domain", is close to the substrate; the other, termed the "AChR domain", is further from the substrate and is rich in acetylcholine re...
journal_title:Experimental cell research
pub_type: 杂志文章
doi:10.1016/0014-4827(89)90261-9
更新日期:1989-06-01 00:00:00