Neurotransplantation in mice: the concorde-like position ensures minimal cell leakage and widespread distribution of cells transplanted into the cisterna magna

Abstract

The access of transplanted cells to large areas of the CNS is of critical value for cell therapy of chronic diseases associated with widespread neurodegeneration. Intrathecal cell application can match this requirement. Here we describe an efficient method for cell injection into the cisterna magna and the assessment of the cell distribution within subarachnoidal space in mice. In order to maximize cell distribution we applied a "concord-like" position, where the cisterna magna is nearly the highest point of the animal's body. A drop of saline on the needle insertion site avoided the outflow of transplanted cells from subarachnoidal space with CSF during surgery. Twenty-four hours later the preparation of the CNS with an intact dura mater by a suitable dissection technique (described in detail) revealed approx. 80% of the injected cells (100,000 cells per animal) within the subarachnoidal space ranging from the skull base (olfactory nerve to premedullary cisterns) to the IV ventricle, and to both the ventral and dorsal surfaces of the spinal cord. Thus the "concorde-like" position proved to be very useful for intrathecal cell application leading to a widespread cell distribution within the subarachnoidal space.