Suitability of allogeneic sertoli cells for ex vivo gene delivery in the injured spinal cord.
Cell-based gene delivery for gene therapy offers the advantages of long-term stable expression of proteins without the safety concerns associated with viral vectors. However, issues of immune rejection prevent the widespread use of allogeneic cell implants. In this study, we determine if Sertoli cells, known for their immune privileged status, are suitable vehicles for allogeneic cell-based gene delivery into the injured spinal cord. As proof of concept, Sertoli cells were modified with recombinant adenovirus expressing enhanced green fluorescent protein (eGFP) or a human trophic factor, neurotrophin-3 (hNT-3), and eGFP. Genetically modified Sertoli cells retained their immunosuppressive ability in vitro, based upon lymphocyte proliferation assays, and were capable of generating biologically relevant levels of NT-3. Similarly, modified, allogeneic cells, implanted into the acutely injured spinal cord, reduced the early inflammatory response while producing significant levels of hNT-3 for at least 3 days after grafting. Moreover, these cells survived for at least 42 days after implantation in the injured cord. Together, these results demonstrate that Sertoli cells function in immunomodulation, can be engineered to produce bioactive molecules, and show long-term survival after implantation into the hostile environment of the acutely injured spinal cord. Such long-term survival represents an important first step toward developing an optimal cell-based delivery system that generates sustained expression of a therapeutic molecule.
Trivedi
Alpa A
AA
MandalMed, Inc., 2645 Ocean Avenue, Suite 302, San Francisco, CA 94132, USA. alpa@mandalmed.com
Igarashi
Takuji
T
Compagnone
Nathalie
N
Fan
Xiaoqing
X
Hsu
Jung-Yu C
JY
Hall
Deborah E
DE
John
Constance M
CM
Noble-Haeusslein
Linda J
LJ
D000818:Animals; D039481:CD11b Antigen / Q000378:metabolism; D049109:Cell Proliferation; D002478:Cells, Cultured; D004195:Disease Models, Animal; D004622:Embryo, Mammalian; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005786:Gene Expression Regulation / Q000502:physiology; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008214:Lymphocytes / Q000502:physiology; D008297:Male; D051379:Mice; D009416:Nerve Regeneration / Q000502:physiology*; D020933:Neurotrophin 3 / Q000235:genetics / Q000502:physiology*; D012708:Sertoli Cells / Q000502:physiology*; D013119:Spinal Cord Injuries / Q000150:complications / Q000628:therapy*; D013997:Time Factors; D014184:Transplantation, Homologous / Q000379:methods
D039481:CD11b Antigen; D020933:Neurotrophin 3; D049452:Green Fluorescent Proteins