Exploration And Research Of A New Approach Of Stem Cell Transplantation For Inner Ear Through The Subarachnoid Cavity

Objectives:To culture and to identify the human umbilical cord mesenchymal stem cells and human induced pluripotent stem cells. To compare two kinds of cells and to discuss whether they are suitable for quickproliferation and to meet the needs of in vivo transplantation.Methods:The human umbilical cord mesenchymal stem cells and induced pluripotent stem cells (iPSc) were cultured and identified. The growth curves were curved and the cell morphologies were compared.Result:The phenotype of mesenchymal stem cells cultured was CD 73 (+), CD 105 (+), CD 31 (-), CD 34 (-), which was in accordance with the characteristics of umbilical cord mesenchymal stem cells. The phenotype of Induced pluripotent stem cells cultured was (NANOG+), OCT-4 (+), SOX-2 (+), SSEA-4 (+), TRA-60 (+) and TRA-81 (+), which was in accordance with the characteristics of Induced pluripotent stem cells. Umbilical cord mesenchymal stem cells were spindle-like and its doubling time was 42 hours. The shape of Induced pluripotent stem cells was round or oval, and the doubling time was 18 hours.Conclusion:Human umbilical cord mesenchymal stem cells and Induced pluripotent stem cells had high proliferation speed and steady state, both of which are suitable for transplantation.Objectives:To research the feasibility of a new approach for stem cell transplantation to inner ear, and to examine the distribution of the induced pluripotent stem cells transplanted through the subarachnoid cavity and the effects on auditory brainstem response (ABR).Methods:The induced pluripotent stem cells (iPSc) were cultured and identified. The induced pluripotent stem cells were transplanted into young congenital deaf albino Rongchang swine through the subarachnoid cavity. And swine with normal hearing injected only physiological saline was used as a control group. Observation window (OBW) was set as 1 day post-operation,3 days post-operation and 7 days post-operation. ABR were tested before and up to OBW after iPSc transplantation. The samples was collected according to the OBW. Immunohistochemistry was used to detect the proliferated cell nuclear antigen (PCNA) from frozen tissue of cochlea. Western Blot was used to detect the protein expressed in the samples and RT-PCR detect the gene expression.Result:The transplanted iPSc was detected inthe inner ear at 3 days and 7 days-post transplantation through Immunohistochemistry. The protein of PCNA from iPSc was found in the spinal cord, medulla, cerebrum, cerebellum, heart, liver, spleen, kidney and lung at the time of OBW through Western Blot. The CETP gene was found expressed in the samples. No wave was detected though the ABR of the young congenital deaf Rongchang swine before and after the transplantation.Conclusion:At the time of OBW, the iPSc could immigrate into inner ears, central nervous system and periphery organs. No wave was detected though the ABR before and after the transplantation with the stimulou intensity more than 120 dB SPL.No further worsening condiction was detected from the spiral ganglion cell.It is a new approach of stem cell transplantation to inner ear through intra-subarachnoid cavity injection and it needs a further exploration for whether it relieve the deafness or not.