title: The use of granulocyte-colony stimulating factor induced mobilization for isolation of dental pulp stem cells with high regenerative potential.
abstract: Human dental pulp stem cells (DPSCs) contain subsets of progenitor/stem cells with high angiogenic, neurogenic and regenerative potential useful for cell therapy. It is essential to develop a safe and efficacious method to isolate the clinical-grade DPSCs subsets from a small amount of pulp tissue without using conventional flow cytometry. Thus, a method for isolation of DPSCs subsets based on their migratory response to optimized concentration of 100 ng/ml of granulocyte-colony stimulating factor (G-CSF) was determined in this study. The DPSCs mobilized by G-CSF (MDPSCs) were enriched for CD105, C-X-C chemokine receptor type 4 (CXCR-4) and G-CSF receptor (G-CSFR) positive cells, demonstrating stem cell properties including high proliferation rate and stability. The absence of abnormalities/aberrations in karyotype and lack of tumor formation after transplantation in an immunodeficient mouse were demonstrated. The conditioned medium of MDPSCs exhibited anti-apoptotic activity, enhanced migration and immunomodulatory properties. Furthermore, transplantation of MDPSCs accelerated vasculogenesis in an ischemic hindlimb model and augmented regenerated pulp tissue in an ectopic tooth root model compared to that of colony-derived DPSCs, indicating higher regenerative potential of MDPSCs. In conclusion, this isolation method for DPSCs subsets is safe and efficacious, having utility for potential clinical applications to autologous cell transplantation.
authors.affiliation: Department of Dental Regenerative Medicine, Center of Advanced Medicine for Dental and Oral Diseases, National Center for Geriatrics and Gerontology, Research Institute, 35 Gengo, Morioka, Obu, Aichi 474-8511, Japan.
authors.forename: Masashi
authors.initials: M
authors.lastname: Murakami
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Horibe
authors.forename: Koichiro
authors.initials: K
authors.lastname: Iohara
authors.forename: Yuki
authors.initials: Y
authors.lastname: Hayashi
authors.forename: Yohei
authors.initials: Y
authors.lastname: Osako
authors.forename: Yoshifumi
authors.initials: Y
authors.lastname: Takei
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Nakata
authors.forename: Noboru
authors.initials: N
authors.lastname: Motoyama
authors.forename: Kenichi
authors.initials: K
authors.lastname: Kurita
authors.forename: Misako
authors.initials: M
authors.lastname: Nakashima
mesh_terms: D000293:Adolescent; D000328:Adult; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D063646:Carcinogenesis / Q000473:pathology; D002454:Cell Differentiation / Q000187:drug effects; D019070:Cell Lineage / Q000187:drug effects; D002469:Cell Separation / Q000379:methods*; D003782:Dental Pulp / Q000166:cytology*; D005434:Flow Cytometry; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology*; D019650:Hematopoietic Stem Cell Mobilization; D006614:Hindlimb / Q000098:blood supply / Q000473:pathology; D006801:Humans; D007511:Ischemia / Q000473:pathology; D051379:Mice; D016513:Mice, SCID; D041681:NIH 3T3 Cells; D018919:Neovascularization, Physiologic / Q000187:drug effects; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012038:Regeneration / Q000187:drug effects*; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D014092:Tooth Root / Q000187:drug effects / Q000502:physiology; D055815:Young Adult
chemical_list: D015415:Biomarkers; D012333:RNA, Messenger; D016179:Granulocyte Colony-Stimulating Factor
keywords: Angiogenesis/vasculogenesis; DPSCs; Dental pulp stem/progenitor cells; G-CSF; Granulocyte-colony stimulating factor (G-CSF); Migration; Pulp regeneration; RT-PCR; Trophic effect; dental pulp stem cells; granulocyte-colony stimulating factor; reverse transcription-polymerase chain reaction