COMPARING SURVIVAL OF PUMA-DEFICIENT & WILDTYPE MSCS TO APOPTOTIC STIMULI FOR AUGMENTED REGENERATION
A.B. Lucas S.S.
In this project, a novel research has been performed to determine the conditions necessary for a successful stem cell transplant using Mesenchymal Stem Cells (MSCs) which are being considered for the treatment of spinal cord and brain injuries. Stem cells tend to die (undergo apoptosis) upon transplantation into the harsh environment of diseased or injured tissue, thereby limiting success of stem cell based therapies. A major pro-apoptotic (death causing) protein PUMA is activated in stem cells post transplantation, in response to a number of cell death stimuli including oxidative stress, protein misfolding and anoxia (oxygen deprivation). In this project, apoptotic rates of PUMA-proficient and PUMA-deficient MSCs derived from mice bone marrow were investigated. Above MSC cultures were incubated for varying time periods under normoxia and anoxia; presence and absence of glucose; and exposure to three different types of apoptotic stimuli CPT, TBH and TUN. Extent of apoptosis was then assessed by examining cell/nuclear morphology after Hoechst staining. The following conclusions are made: i) Apoptosis increases with incubation time, deprivation of oxygen, deficiency of glucose and presence of apoptotic stimuli, ii) Anoxia and TBH are strong inducers of apoptosis, iii) Lack of glucose induces apoptosis but not substantially, iv) PUMA-deficient MSCs are more resistant to apoptosis than PUMA-proficient MSCs, under all the circumstances stated in i). Based on this novel study, measures can now be undertaken to remove the pro-apoptotic PUMA gene before transplant of MSCs thereby making stem cell transplants more successful in treating critical spinal cord and brain injuries.
|Second Award – Medicine and Health Sciences – Presented by National Institutes of Health||$1500|