Human brain subventricular zone-derived neural stem cells (NSCs) isolation from stroke patients – A pilot study in hospital University Sains Malaysia (HUSM)

Abstract

Ischemic stroke is caused by cerebral vessel occlusion, resulting in an area of ischemia where the brain tissue irreversibly damaged due to inadequate oxygen and nutrients. Neural stem cell (NSC) grafting has emerged as a potential regenerative therapy for stroke. Here, we described a pilot clinical study of NSC isolation from subventricular zone (SVZ) of human brain. In this preliminary study, nine patients admitted to Hospital University Sains Malaysia (HUSM) with cerebral vascular accident (CVA, i.e. ischemic or hemorrhagic stroke) and necessitating brain resection were recruited. During surgery, SVZ biopsies were excised, weighted and the morphological structure of clinical biopsies were recorded. Then the biopsies were digested and cultured using neurosphere or monolayer culture methods. Out of nine clinical samples, we successfully obtained neurospheres-like cells from five of the patient’s SVZ biopsy, while monolayer culture grew out from two patients’ SVZ biopsy. Characterization assay using immunocyto-chemistry probed with nestin (NSC marker) successfully proved that these cells were NSCs. For the rest of samples, the isolation of SVZ derived-NSC was unsuccessful due to the insufficient SVZ tissue sampling. The tissue obtained was mainly connective or blood tissue, proved by the accumulation of floating cells after 24 hours in culture. In conclusion, NSCs were successfully obtained from some stroke patients SVZ. However, the quantity and quality of isolated cells are tissue-dependent, thus, surgical sampling technique and biopsy sizes are important in ensuring a good NSC isolation efficiency. This study could be an essential guidance for future NSC isolation study in HUSM.