Introduction: Regeneration of tissues in human central nervous system (CNS) is very poor and cell damage can prolong the neurological deficit after CNS injury. For this rationale, considerable attention has focused to generate the large homogenous populations of neural precursor cells (NPCs) in vitro for investigating their possible applications in cell replacement therapy of the damaged CNS. Owing to this various significant developments and modifications have been made in neural stem cell research which is needed to be more specified and enrolment in clinical studies using advanced approaches. The present study was designed to identify the various cell population in human fetal sub ventricular zone (SVZ) derived cells. The study was also focused to know the in vitro behavior and characteristics of NPCs of different gestational age fetus.
Methods: Human fetuses (gestation between 18-21 weeks) were used to isolate the SVZ derived cells. Different cell populations were identified by specific markers such as Nestin, Sox-2, Oct-4, Notch-2, ABCB1, ABCG2, ß-tubulin III, GFAP and O4 using immunocytochemical staining, flow cytometry and molecular analysis. NPCs were magnetically sorted using CD133 and maintained in suspension culture for 21 days.
Results: SVZ derived cells showed positivity for all selected markers except O4. The expression intensity of CD133 and Nestin were found high in 20 gestation period. FDA analysis showed >92% viability in CD133+ cells. In suspension culture =100µm diameter neurospheres were formed after 14 days and able to survive for 21 days of initial plating. Nestin staining showed positive result for morphological characterization of neurospheres.
Conclusions: The present study revealed that key cell population in SVZ is NPCs though it also contains mature dedicated neuronal and astrocyte cells. In vitro expanded NPCs show an option to resolve the paucity of NPCs accessibility for their use in the treatment of CNS diseases.
Patient Care: In vitro expanded NPCs show an option to resolve the paucity of NPCs accessibility for their use in the treatment of CNS diseases.
Learning Objectives: exploring the possibility of across the counter cell therapy products in neural regeneration