Ex vivo and in vitro studies of transgene expression in rat astrocytes transduced with lentiviral vectors.

• Implantation of cells genetically modified to express therapeutic genes into the brain has been proposed as a potential treatment for neurodegenerative diseases. In the current study embryonic rat-derived astrocytes were cultured and transduced with a lentiviral vector expressing the reporter gene green fluorescent protein (GFP) and subsequently grafted into the adult rat brain. The proportion of GFP expressing cells was stable, albeit small (1%), at all survival times, up to 6 weeks, the longest time point studied. In parallel in vitro studies, the astrocytes were lentivirally transduced to express either one of the two isoforms of glutamate decarboxylase (GAD(65) or GAD(67)) or glial cell line-derived neurotrophic factor (GDNF). When transducing 293T cells with the two GAD vectors, released GABA could be measured using high-performance liquid chromatography. Further studies of rat astrocytes transduced with the same vectors resulted in a level of GAD activity about 10 times higher than the activity of an intact rat striatum. One hundred thousand astrocytes transduced with LV-GDNF released approximately 27 ng of GDNF per hour. Thus, taken together, our observations provide support for the use of rat astrocytes in ex vivo gene transfer of these proteins in animal models of CNS disorders, e.g., Parkinson's disease or epilepsy.

Ämnesord

MEDICIN OCH HÄLSOVETENSKAP  -- Klinisk medicin -- Neurologi (hsv//swe)

MEDICAL AND HEALTH SCIENCES  -- Clinical Medicine -- Neurology (hsv//eng)

Nyckelord

Gene Expression : physiology

gamma-Aminobutyric Acid : metabolism

Transgenes : physiology

Transduction Genetic

Support Non-U.S. Gov't

Rats Sprague-Dawley

Rats

Luminescent Proteins : biosynthesis : genetics

Lentivirus : genetics

Kidney : metabolism

Immunohistochemistry

Human

Genes Reporter

Genetic Vectors : genetics : metabolism

Corpus Striatum : cytology : metabolism

Cells Cultured

Brain Tissue Transplantation

Astrocytes : cytology : metabolism : transplantation

Animal

Publikations- och innehållstyp

art (ämneskategori)

ref (ämneskategori)