Hypoxic-ischemic brain damage is one of the main causes of death and neurological disability after stroke worldwide. Our previous studies have demonstrated that interleukin-1 (IL-1) primed mesenchymal stem cells (MSCs) have a stronger anti-inflammatory and neurotrophic phenotype and can be used as a new cellular stroke therapy. In this project, we will study the effects of IL-1 primed MSCs on oxygen and glucose deprivation (OGD) injury in neurons and astrocytes, with the ultimate aim to explore the potential clinical application of IL-1 primed MSCs on neuroprotection.
The project will have three main objectives:
1) To test the effects of IL-1 primed MSCs on neuroprotection after experimental cerebral ischemia
Adults mice exposed to hypoxia/ischemia will be injected with IL-1-primed MSCs. Field excitatory postsynaptic potentials in the hippocampal CA1 region of brain slices will be recorded to examine the effects of primed MSC transplantation on hippocampal synaptic plasticity. The effect of MSCs on hypoxia/ischemia brain injury, inflammation and repair will be performed at various times post MSCs injections by routine histology, immunohistochemistry and ELISAs.
2) To test the effects of IL-1 primed MSCs on OGD-induced neuronal injury.
Primary culture of hippocampal neurons will be subjected to OGD-induced injury and subsequently treated with primed MSCs. Western blot will be used to detect apoptotic markers and ELISA will be used to detect the expression of neuronal-derived cytokines.
3) To test the effects of IL-1 primed MSCs on OGD-induced astrocytic injury.
Primary astrocytes will used in similar experiments to those described for neurones above. Apoptosis and astrogliosis will be assessed by immunocytochemistry and Western blot using established markers.
Training/techniques to be provided:
The student will be provided with all the techniques that are required to achieve the aims of the projects, including in vivo models of hypoxia / ischemia, histology, preclinical brain imaging, analysis of key markers of tissue injury / repair by immunohistochemistry / ELISA. Furthermore, full training will be provided to successfully conduct all in vitro experiments, including cell cultures, organotypic cultures, oxygen glucose deprivation and a large array of analytical techniques.
Candidates are expected to hold (or be about to obtain) a minimum upper second class honours degree (or equivalent) in a related area / subject. Candidates with experience in stem cell biology/therapy or with an interest in regenerative medicine in brain diseases are encouraged to apply.
For international students we also offer a unique 4 year PhD programme that gives you the opportunity to undertake an accredited Teaching Certificate whilst carrying out an independent research project across a range of biological, medical and health sciences. For more information please visit http://www.internationalphd.manchester.ac.uk
Applications are invited from self-funded students. This project has a Band 2 fee. Details of our different fee bands can be found on our website (View Website). For information on how to apply for this project, please visit the Faculty of Biology, Medicine and Health Doctoral Academy website (View Website).
As an equal opportunities institution we welcome applicants from all sections of the community regardless of gender, ethnicity, disability, sexual orientation and transgender status. All appointments are made on merit.
Wong R, Lénárt N, Hill L, Toms L, Coutts G, Martinecz B, Császár E, Nyiri G, Papaemmanouil A, Waisman A, Müller W, Schwaninger M, Rothwell N, Francis S, Pinteaux E*, Denés A*, Allan SM* (2018) Interleukin-1 mediates ischaemic brain injury via distinct actions on endothelial cells and cholinergic neurons. Brain Behav Immun. 2018 Nov 16. pii: S0889-1591(18)30403-3. doi: 10.1016/j.bbi.2018.11.012. * joint senior authors
Rajkovic I, Wong R, Lemarchand E, Rivers-Auty J, Rajkovic O, Garlanda C, Allan SM, Pinteaux E. (2018) Pentraxin 3 promotes long-term cerebral blood flow recovery, angiogenesis and neuronal survival after stroke. J Mol Med. 96, 1319-1332. doi: 10.1007/s00109-018-1698-6
Redondo-Castro E, Cunningham C, Miller J, Cain SA, Allan SM, Pinteaux E (2018) Generation of human mesenchymal stem cell 3D spheroids using low-binding plates. Bioprotoc. 20;8(16). pii: e2968. doi: 10.21769/BioProtoc.2968.
Redondo-Castro E, Cunningham C, Miller J, Brown H, Allan SM, Pinteaux E. (2018) Changes in the secretome of tri-dimensional spheroid-cultured human mesenchymal stem cells in vitro by interleukin-1 priming. Stem Cell Res Ther. 9,11.
Redondo-Castro E, Cunningham C, Miller J, Martuscelli L, Aoulad-Ali S, Rothwell NJ,Kielty CM, Allan SM, Pinteaux E (2017) Interleukin-1 primes human mesenchymal stem cells towards an anti-inflammatory and pro-trophic phenotype in vitro. Stem Cell Res Ther. 8, 79.