Abstract
In our lab we study neurogenesis and the development of brain tumors. We work towards treatment strategies for glioblastoma and towards using autologous neural stem cells for tissue regeneration strategies for brain damage and neurodegenerative disorders. It has been our policy to try to establish living cell cultures from all human biopsy material that we obtain. We hypothesized that small pieces of brain tissue could be cryopreserved and that live neural stem cells could be recovered at a later time. DMSO has been shown to possess a remarkable ability to diffuse through cell membranes and pass into cell interiors. Its chemical properties prevent the formation of damaging ice crystals thus allowing cell storage at or below −180 C. We report here a protocol for successful freezing of small pieces of tissue derived from human brain and human brain tumours. Virtually all specimens could be successfully revived. Assays of phenotype and behaviour show that the cell cultures derived were equivalent to those cultures previously derived from fresh tissue.
Highlights
In our lab we study neurogenesis and the development of brain tumors
Types of tissue samples were subventricular zone (SVZ), Hippocampus (HPC), Cortex (Grey and White Matter), Grey Matter (GM), and White Matter (WM) The stem cell we described has the ability to generate vast numbers of cells, differentiate into neurons, astrocytes and oligodendrocytes
Biopsies to be cryogenically frozen were able to vary in size between 100 mm[3] down to 8 mm3. 100 mm[3] (100 μL) was the maximum quantity able to be stored in a 1 mL cryotube containing 100 μL dimethyl sulphoxide (DMSO) and the remaining volume to a total of 1 mL with serum and thence subsequently be able to produce viable cells efficiently
Summary
In our lab we study neurogenesis and the development of brain tumors. We work towards treatment strategies for glioblastoma and towards using autologous neural stem cells for tissue regeneration strategies for brain damage and neurodegenerative disorders. It has been our policy to try to establish living cell cultures from all human biopsy material that we obtain. We hypothesized that small pieces of brain tissue could be cryopreserved and that live neural stem cells could be recovered at a later time. In 2013 we published a comprehensive study of neural stem cells derived from human brain biopsies[9]. For the first time multipotency of adult human brain-derived stem cells was demonstrated beyond tissue boundaries. In 1979, Clafrin and Malinin reported cryopreservation of small pieces of tissue from various human organs and subsequent thawing and culture of live cells[11]. We hypothesized that small pieces of brain tissue could be cryopreserved and that live neural stem cells could be recovered at a later
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