Asian Spine J.  2017 Dec;11(6):870-879. 10.4184/asj.2017.11.6.870.

Generation of Induced Pluripotent Stem Cells and Neural Stem/Progenitor Cells from Newborns with Spina Bifida Aperta

Affiliations
  • 1Department of Physiology, Keio University School of Medicine, Tokyo, Japan. ybamba@osakah.johas.go.jp
  • 2Department of Pediatric Neurosurgery, Kansai Medical University, Hirakata, Japan.
  • 3Department of Pediatric Neurosurgery, National Medical Center for Children and Mothers Research Institute, Tokyo, Japan.
  • 4Division of Stem Cell Research, Institute for Clinical Research, Osaka National Hospital, National Hospital Organization, Osaka, Japan.
  • 5Division of Regenerative Medicine, Institute for Clinical Research, Osaka National Hospital, National Hospital Organization, Osaka, Japan.
  • 6Laboratory Animal Research Department, Biomedical Research Laboratory, Central Institute for Experimental Animals, Kawasaki, Japan.
  • 7CRIFM Clinical Research Institute of Fetal Medicine Pooh Maternity Clinic, Osaka, Japan.
  • 8Department of Neurosurgery, Osaka National Hospital, National Hospital Organization, Osaka, Japan.
  • 9Department of Pediatric Neurosurgery, Takatsuki General Hospital, Takatsuki, Japan.

Abstract

STUDY DESIGN: We established induced pluripotent stem cells (iPSCs) and neural stem/progenitor cells (NSPCs) from three newborns with spina bifida aperta (SBa) using clinically practical methods. PURPOSE: We aimed to develop stem cell lines derived from newborns with SBa for future therapeutic use. OVERVIEW OF LITERATURE: SBa is a common congenital spinal cord abnormality that causes defects in neurological and urological functions. Stem cell transplantation therapies are predicted to provide beneficial effects for patients with SBa. However, the availability of appropriate cell sources is inadequate for clinical use because of their limited accessibility and expandability, as well as ethical issues.
METHODS
Fibroblast cultures were established from small fragments of skin obtained from newborns with SBa during SBa repair surgery. The cultured cells were transfected with episomal plasmid vectors encoding reprogramming factors necessary for generating iPSCs. These cells were then differentiated into NSPCs by chemical compound treatment, and NSPCs were expanded using neurosphere technology.
RESULTS
We successfully generated iPSC lines from the neonatal dermal fibroblasts of three newborns with SBa. We confirmed that these lines exhibited the characteristics of human pluripotent stem cells. We successfully generated NSPCs from all SBa newborn-derived iPSCs with a combination of neural induction and neurosphere technology.
CONCLUSIONS
We successfully generated iPSCs and iPSC-NSPCs from surgical samples obtained from newborns with SBa with the goal of future clinical use in patients with SBa.

Keyword

Myelomeningocele; Spina bifida; Induced pluripotent stem cells; Neural stem/progenitor cells; Regenerative medicine

MeSH Terms

Cells, Cultured
Ethics
Fibroblasts
Humans
Induced Pluripotent Stem Cells*
Infant, Newborn*
Meningomyelocele
Plasmids
Pluripotent Stem Cells
Regenerative Medicine
Skin
Spina Bifida Cystica*
Spinal Cord
Spinal Dysraphism*
Stem Cell Transplantation
Stem Cells
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