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Strongly amphiphilic photosensitizers are not substrates of the cancer stem cell marker ABCG2 and provides specific and efficient light-triggered drug delivery of an EGFR-targeted cytotoxic drug.
Selbo PK, Weyergang A, Eng MS, Bostad M, Maelandsmo GM, Hogset A, Berg K (2012) Strongly amphiphilic photosensitizers are not substrates of the cancer stem cell marker ABCG2 and provides specific and efficient light-triggered drug delivery of an EGFR-targeted cytotoxic drug. J Control Release 159(2):197-203. doi: 10.1016/j.jconrel.2012.02.003
Summary: Many anti-cancer drugs are substrates of the ATP-binding cassette transporter ABCG2. Unfortunately ABCG2 is also thought to play an important role in multi-drug resistance and the protection of cancer stem cells against chemotherapeutics and photodynamic therapy. This paper examined whether photosensitizers used in photochemical internalization (PCI) are substrates for ABCG2. Streptavidin-ZAP (Cat. #IT-27) was combined with biotinylated EGF and applied to cells in culture; saporin (Cat. #PR-01) was used as a control. The data show that PCI with the EGF-saporin toxin did not utilize ABCG2 to enter cells.
Related Products: Streptavidin-ZAP (Cat. #IT-27), Saporin (Cat. #PR-01)
Novel application of human periodontal ligament stem cells and water-soluble chitin for collagen tissue regeneration: in vitro and in vivo investigations.
Jung IH, Park JC, Kim JC, Jeon DW, Choi SH, Cho KS, Im GI, Kim BS, Kim CS (2012) Novel application of human periodontal ligament stem cells and water-soluble chitin for collagen tissue regeneration: in vitro and in vivo investigations. Tissue Eng Part A 18(5-6):643-53. doi: 10.1089/ten.TEA.2011.0164 PMID: 21981356
Usage: Western blot 1:1000
Related Products: Trans-4-Hydroxy-L-Proline Rabbit Polyclonal, Conjugated (Cat. #AB-T044)
Generation of human melanocytes from induced pluripotent stem cells
Ohta S, Imaizumi Y, Okada Y, Akamatsu W, Kuwahara R, Ohyama M, Amagai M, Matsuzaki Y, Yamanaka S, Okano H, Kawakami Y (2011) Generation of human melanocytes from induced pluripotent stem cells. PLoS One 6(1):e16182. doi: 10.1371/journal.pone.0016182 PMID: 21249204
Summary: For immunocytochemistry, cells were fixed with PBS containing 4% PFA for 20 min at room temperature. Then, cells were subjected to immunofluorescence staining (1∶100).
Related Products: NGFr (ME20.4, p75) Mouse Monoclonal (Cat. #AB-N07)
Functional cholinergic neurons from human embryonic stem cells
Liu Y, Krencik R, Liu H, Ma L, Zhang X, Zhang S-C (2010) Functional cholinergic neurons from human embryonic stem cells. Neuroscience 2010 Abstracts 331.5/B19. Society for Neuroscience, San Diego, CA.
Summary: Basal forebrain cholinergic neurons play a critical role in regulating memory and cognition. Degeneration or dysfunction of these neurons is associated with neurological conditions including Alzheimer’s disease and dementia. In this study, we aimed at generating cholinergic neurons from human embryonic stem cells (hESCs) for therapeutic development. hESCs were first differentiated to primitive neuroepithelial cells in a chemically defined medium. In the presence of sonic hedgehog, over 97% of the differentiated cells became Nkx2.1-expressing ventral forebrain progenitors. These ventral progenitors further differentiated to cholinergic neurons with basal forebrain characteristics by expressing ChAT, VAChAT, FoxG1, Nkx2.1, Islet1, ßIII-tubulin, MAP2, P75, Synapsin but not GABA, Glutamate, or Mash2. The hESC-generated cholinergic neurons were electrophysiologically active in vitro. Following transplantation into the hippocampus of mice, in which cholinergic neurons in the medial septum were destroyed by IgG-P75-saporin, the grafted human cells produced large cholinergic neurons. The animals transplanted with cholinergic neurons demonstrated an improvement in learning and memory deficit. These results indicate that the human stem cell-generated cholinergic neurons are functional, thus providing a new source for drug discovery and cell therapy for neurological disorders that affect cholinergic neurons.
Related Products: mu p75-SAP (Cat. #IT-16)
The recovery of locomotion after lumbar spinal cord motoneuron depletion is affected by the modulation of Sonic Hedgehog and Notch-1 pathways
Gulino R, Gulisano M (2010) The recovery of locomotion after lumbar spinal cord motoneuron depletion is affected by the modulation of Sonic Hedgehog and Notch-1 pathways. Neuroscience 2010 Abstracts 259.19/W12. Society for Neuroscience, San Diego, CA.
Summary: Sonic hedgehog (Shh) and Notch-1 are involved in the regulation of stem cell function. Additionally, Notch-1 has a role as modulator of synaptic plasticity. In our previous work, we injected Cholera toxin-B saporin (CTB-sap) into the gastrocnemius muscle to induce a selective depletion of motoneurons within lumbar mice spinal cord (SC) and analysed the expression levels of Shh, Notch-1, Numb, Choline acetyltransferase (ChAT) and Synapsin-I proteins. The functional outcome of the lesion was monitored by grid walk test and rotarod. Shh and Notch-1 appeared reduced in the lesioned tissue and correlated with ChAT and Synapsin-I levels, suggesting a role in modulating synaptic plasticity. Numb expression was also reduced after lesion and appeared correlated with motor performance but not with synaptic plasticity. We sought to determine if the pharmacological manipulation of the expression of Shh and Notch-1 could affect functional recovery by a mechanism involving synaptic plasticity. Therefore, we used CTB-sap to induce lesion as above, and injected Cyclopamine, recombinant Shh or recombinant DLL4 chronically into the intrathecal space by means of osmotic minipumps for two weeks. The functional recovery was monitored for one month after lesion by means of grid walk test, two times a week, whereas the modifications of Shh, Notch-1, ChAT and Synapsin-I protein expression levels were measured by western blot and immunohistochemistry. Here, we show that the modulation of Shh or Notch-1 pathways could affect the recovery of locomotion. Moreover, the molecular mechanisms underlying this process is discussed.
Related Products: CTB-SAP (Cat. #IT-14)
Inhibition of notch signaling in glioblastoma targets cancer stem cells via an endothelial cell intermediate.
Hovinga KE, Shimizu F, Wang R, Panagiotakos G, Van Der Heijden M, Moayedpardazi H, Correia AS, Soulet D, Major T, Menon J, Tabar V (2010) Inhibition of notch signaling in glioblastoma targets cancer stem cells via an endothelial cell intermediate. Stem Cells 28:1019-1029. doi: 10.1002/stem.429 PMID: 20506127
Summary: The antibody CD105 (1:1,000) was incubated with Mab-ZAP, to allow binding and formation of a CD105 antibody-saporin complex, which was added to explants or to a human cerebral microvessel endothelial cell line as control. CD105 antibody was also incubated with a Goat-IgG-SAP for control that does not bind CD105. The conjugates were injected into the explant under a dissecting microscope after gently incising the explant surface for better access.
Related Products: Mab-ZAP (Cat. #IT-04), Goat IgG-SAP (Cat. #IT-19)
Expression of cell fate determinants and plastic changes after neurotoxic lesion of adult mice spinal cord by cholera toxin-B saporin.
Gulino R, Perciavalle V, Gulisano M (2010) Expression of cell fate determinants and plastic changes after neurotoxic lesion of adult mice spinal cord by cholera toxin-B saporin. Eur J Neurosci 31(8):1423-1434. doi: 10.1111/j.1460-9568.2010.07170.x
Summary: Sonic hedgehog, Notch-1, and Numb are proteins known to be involved in the function of stem cells. Understanding of how they might work in adults may provide methods to improve recovery from spinal cord injury. In this work the authors injected 3 µg of CTB-SAP (Cat. #IT-14) into the medial and lateral gastrocnemius muscles of mice. Analysis of protein levels following motoneuron depletion gives some insight into the molecular framework of nerve injury.
Related Products: CTB-SAP (Cat. #IT-14)
Derivation of neural crest cells from human pluripotent stem cells
Lee G, Chambers SM, Tomishima MJ, Studer L (2010) Derivation of neural crest cells from human pluripotent stem cells. Nat Protoc 5(4):688-701. doi: 10.1038/nprot.2010.35 PMID: 20360764
Summary: Protocols are presented for the purification and propagation of hPSC-NC cells using flow cytometry and defined in vitro culture conditions. This protocol has been validated in multiple independent hESC and hiPSC lines. The average time required for generating purified hPSC-NC precursors using this protocol is 2–5 weeks.
Usage: Neural crest cells (1:200).
Related Products: NGFr (ME20.4, p75) Mouse Monoclonal (Cat. #AB-N07)
Poster: Expression of cell fate determinants and plastic changes after neurotoxic lesion of adult mice spinal cord by cholera toxin-B saporin.
Gulino R, Gulisano M (2009) Poster: Expression of cell fate determinants and plastic changes after neurotoxic lesion of adult mice spinal cord by cholera toxin-B saporin. Neuroscience 2009 Abstracts 563.15/DD51. Society for Neuroscience, Chicago, IL.
Summary: Recent studies have attempted to achieve recovery after spinal cord (SC) injury or disease by either increase neurogenesis or stimulate neuroplasticity. Sonic hedgehog (Shh) Notch-1 and Numb are involved in the regulation of stem cell function. Additionally, Notch-1 has a role as modulator of synaptic plasticity. Little is known about the role of these proteins in the adult SC after selective removal of motoneurons. We injected Cholera toxin-B saporin into the gastrocnemius muscle to induce a selective depletion of motoneurons within lumbar mice SC and analysed the expression levels of Shh, Notch-1, Numb, Choline acetyltransferase (ChAT) and Synapsin-I proteins. The functional outcome of the lesion was monitored by grid walk test and rotarod. The neurotoxin lesion determined a motoneuron depletion and a decrease of ChAT and Synapsin-I protein levels in the lumbar SC. ChAT and Synapsin-I appeared correlated each other and with the motor performance, suggesting that the recovery of locomotion could depend from synaptic plasticity. Moreover, we observed a number of proliferating cells within the depleted SC, which were identified as active astrocytes. Shh and Notch-1 appeared reduced in the lesioned tissue and correlated with ChAT and Synapsin-I levels, suggesting a role in modulating synaptic plasticity. Numb expression was also reduced after lesion and appeared correlated with motor performance Therefore, given the role of these proteins in adult neurogenesis, we presume their involvement also in the observed glial reaction. The in vivo manipulation of Shh, Notch-1 and Numb signalling after lesion could be a way to reduce glial reaction and improve functional recovery.
Related Products: CTB-SAP (Cat. #IT-14)
The role of microglia and neuropeptides in regulating hippocampal neurogenesis.
Sivasathiaseelan H, Nunan R, Zaben M, Shtaya A, Gray WP (2009) The role of microglia and neuropeptides in regulating hippocampal neurogenesis. Neuroscience 2009 Abstracts 31.26/B52. Society for Neuroscience, Chicago, IL.
Summary: Adult mammalian neurogenesis is evident in the hippocampal dentate gyrus where it plays a role in learning and memory and is implicated in the pathophysiology of several brain disorders. Microglia, the innate immune cells of the brain, have recently emerged as an important component of the neurogenic niche, however their role in the regulation of neurogenesis under physiological and pathophysiological conditions is a matter of debate. The aim of this study is to investigate the effect of microglia on hippocampal neurogenesis and to look at how vasoactive intestinal peptide (VIP), a potent immunomodulatory neuropeptide found in dentate gyrus interneurons, modulates the effects microglia have on neurogenesis. In this study, we have investigated the effect of microglial depletion (using MAC-SAP), microglial co-culture and addition of microglia-conditioned-medium on primary hippocampal cell cultures derived from post-natal rats. We have also looked at how pre-treatment of microglia with VIP alters their effect on hippocampal cultures. Bromodeoxyuridine was used as a marker of cell proliferation. Quantification of cell death was achieved using the nuclear stain 4′,6-diamidino-2-phenylindole and Propidium Iodide. Immunohistochemistry was used to phenotype cells for nestin, GFAP and Tuj1. We have shown that microglial depletion results in a reduction in the numbers of nestin, GFAP and Tuj1 expressing cells. This reduction has been shown to be attributable to a decrease in cell survival and proliferation. Conversely, co-culture of microglia with hippocampal neurons or addition of their conditioned medium results in increased cell survival and proliferation. Pre-treatment of microglia with VIP was shown to increase both their proliferative and trophic effect on hippocampal cultures. In conclusion, this study demonstrates that microglia induce proliferative and trophic effects on neural stem cells and immature neurons through the release of soluble factors. Furthermore, we provide evidence that VIP regulates the release of these soluble factors, thus identifying a novel neuro-immuno-neurogenic link.
Related Products: Mac-1-SAP rat (Cat. #IT-33)