Development of stem cell based technologies and regenerative medicines
Stem cells have remarkable properties, unlike mature cells that play an important bodily function eg. Liver cells or muscle cells, all stem cells are unspecialised and can keep proliferating and increasing in number indefinitely. Additionally, pluripotent stem cells (PSC) have the capacity to mature into all cell types of the body. Therefore, these PSC have great potential for the development of model systems to study different cell types affected by disease and also present the opportunity to develop cellular or regenerative therapies.
We are interesting in finding new reagents and techniques to maintain these important cells in the laboratory and to efficiently induce them to differentiate into the mature cells that we want to work with. Our specific interests are in blood and blood vessel development, disease and therapies.
Progress towards the cGMP production of hESC derived RBCs
Blood Transfusion has become a mainstay of modern medical practice. However problems persist both nationally and internationally in maintaining adequacy of supply, managing the risk of transmission of infectious agents and immune incompatibility between donor and recipient. Human embryonic and induced pluripotent stem cells (hESCs & iPSC) have unique properties in that they can be maintained indefinitely in culture in an undifferentiated state and yet retain the ability to form all the cells and tissues within the body. They therefore offer a potentially limitless source from which to generate red cells (RBCs) for use in clinical transfusion. Initially in vitro RBCs may have particular utility for patients who receive regular transfusions such as those with haemoglobinopathies. Within the project we have generated hESC to cGMP grade (Roslin Cells Ltd) in compliance with UK regulatory requirements for eventual clinical use. We are able to differentiate these to form haematopoietic progenitor cells (HPC) resulting in ≥95% conversion to erythroid cells with high expansion in cell numbers, in a stromal free, suspension based culture system. We have also demonstrated that this methodology is similarly effective for iPSC.
Funded by the Wellcome Trust, Scottish Funding Council and SNBTS
Generation of vascular endothelial cells from pluripotent stem cells.
Our work is based on the refinement and application of vascular endothelial cell production for regenerative medicine within the context of ischemic cardiovascular diseases. Our principal focus is on the basic biology of differentiation to vascular endothelium and the application of this to in vivo peripheral and myocardial vasculogenesis. This area has received significantly less attention than the production of cardiomyocytes and yet offers the potential for therapy for a large population of patients if effectively harnessed. Our proposed programme of work is founded on collective data demonstrating our ability to induce differentiation of hES cells to vascular endothelial cells in a highly efficient feeder-free, serum-free manner with potential for scale-up, our ability to reprogramme somatic cells to iPS cells from patients with cardiovascular disease. Our aim is to fully dissect key signalling events, transcriptional networks and epigenetic changes that lead to effective hESC and iPSC-derived endothelial cells with assessment of vasculogenesis in vivo and, ultimately, in patients.
Funded by the British Heart Foundation, MRC and EPSRC
- Kaupisch A, Kennedy L, Stelmanis V, Tye B, Kane NM, Mountford JC, Courtney A, Baker AH. Derivation of Vascular Endothelial Cells from Human Embryonic Stem Cells Under GMP-Compliant Conditions: Towards Clinical Studies in Ischaemic Disease. J Cardiovasc Transl Res 2012;5:605-617
- Kane NM, Howard L, Descamps B, Meloni M, McClure JD, Lu R, McCahill A, Breen C, Mackenzie RM, Delles C, Mountford JC, Milligan G, Emanueli C, Baker AH. A Role for microRNAs 99b, 181a and 181b in the Differentiation to Vascular Endothelial Cells from Human Embryonic Stem Cells. Stem Cells 2012;30:643-654
- Marenah L, Allan EK, Mountford JC, Holyoake TL, Jørgensen HG, Elliott MA. Investigation into omacetaxine solution stability for in vitro study. Biomed Chromatogr 2012;26:545-547
- Yung S, Ledran M, Moreno-Gimeno I, Conesa A, Montaner D, Dopazo J, Dimmick I, Slater NJ, Marenah L, Real PJ, Paraskevopoulou I, Bisbal V, Burks D, Santibanez-Koref M, Moreno R, Mountford J, Menendez P, Armstrong L, Lako M. Large-scale transcriptional profiling and functional assays reveal important roles for Rho-GTPase signalling and SCL during haematopoietic differentiation of human embryonic stem cells. Hum Mol Genet 2011;20:4932-4936
- Kane NM, Nowrouzi A, Mukherjee S, Blundell MP, Greig JA, Lee WK, Houslay MD, Milligan G, Mountford JC, von Kalle C, Schmidt M, Thrasher AJ, Baker AH. Lentivirus-mediated reprogramming of somatic cells in the absence of transgenic transcription factors. Mol Ther 2010;12:2139-2145
- Burton P, Adams DR, Abraham A, Allcock RW, Jiang Z, McCahill A, Gilmour J, McAbney J, Kaupisch A, Kane NM, Baillie GS, Baker AH, Milligan G, Houslay MD, Mountford JC. Erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA) blocks differentiation and maintains the expression of pluripotency markers in human embryonic stem cells. Biochem J 2010;432:575-584
- Andrews PD, Becroft M, Aspegren A, Gilmour J, James MJ, McRae S, Kime R, Allcock RW, Abraham A, Jiang Z, Strehl R, Mountford JC, Milligan G, Houslay MD, Adams DR, Frearson JA. High-content screening of feeder-free human embryonic stem cells to identify pro-survival small molecules. Biochem J 2010;432:21-33
- Kane NM, Meloni M, Spencer HL, Craig MA, Strehl R, Milligan G, Houslay MD, Mountford JC, Emanueli C, Baker AH. Derivation of endothelial cells from human embryonic stem cells by directed differentiation: analysis of microRNA and angiogenesis in vitro and in vivo. Arterioscler Thromb Vasc Biol 2010;30:1389-1397
- Khanim FL, Hayden RE, Birtwistle J, Lodi A, Tiziani S, Davies NJ, Ride JP, Viant MR, Gunther UL, Mountford JC, Schrewe H, Green RM, Murray JA, Drayson MT, Bunce CM. Combined bezafibrate and medroxyprogesterone acetate: potential novel terapy for acute myeloid leukaemia. PLoS One, 2009;4:e8147
- Pellicano F, Copland M, Jorgensen HJ, Mountford JC, Leber B and Holyoake TL. BMS-214662 induces mitochondrial apoptosis in CML stem/progenitor cells, including CD34+38- cells, through activation of protein kinase C . Blood 2009;114:4186-96
- Davies A, Jordanides NE, Giannoudis A, Lucas CM, Hatziieremia S, Harris RJ, Jørgensen HG, Holyoake TL, Pirmohamed M, Clark RE and Mountford JC. Mountford JC. Nilotinib concentration in cell lines and primary CD34+ chronic myeloid leukemia cells is not mediated by active uptake or efflux by major drug transporters. Leukaemia 2009;23:1999-2006
- Hatziieremia S, Jordanides NE, Holyoake TL, Jørgensen HG and Mountford JC. Inhibition of MDR1 does not sensitise primitive chronic myeloid leukaemia CD34+ cells to imatinib. Exp Hematol 2009;7:692-700
- Myssina S, Helgason GV, Serrels A, Jørgensen HG, Bhatia R, Modi H, Helgason GV, Mountford JC. Hamilton A, Schemionek M, Koschmieder S, Brunton V, Holyoake TL. Combined BCR-ABL inhibition with lentiviral-delivered shRNA and dasatinib augments induction of apoptosis in Philadelphia positive cells. Exp Hematol 2009;37:206-14
- Mountford JC, Turner M. In vitro production of red blood cells. Transfus Apher Sci 2011;45:85-89
- Burton P, Adams DR, Abraham A, Allcock RW, Jiang Z, McCahill A, Gilmour J, McAbney J, Kane NM, Baillie GS, McKenzie FR, Baker AH, Houslay MD, Mountford JC, Milligan G. Identification and characterization of small-molecule ligands that maintain pluripotency of human embryonic stem cells. Biochem Soc Trans 2010;38:1058-1061
- Mountford J, Olivier E, Turner M. Prospects for the manufacture of red cells for transfusion. Br J Haematol 2010;149:22-34
- Mountford JC, Olivier E, Jordanides NE, de Sousa P, Turner ML. Red blood cellsfrom pluripotent stem cells for use in transfusion. Regen Med 2010;5:411-23