Your browser doesn't support javascript.
loading
Method for selective quantification of adipose-derived stromal/stem cells in tissue.
Nishimura, Akira; Kumagai, Takeo; Nakatani, Masaru; Yoshimura, Kotaro.
Afiliación
  • Nishimura A; Kaneka Corporation, Kobe MI R&D Center F 6-7-3, Minatojima, Minamimachi, Chuo-ku, Kobe, Hyogo 650-0047, Japan.
  • Kumagai T; Kaneka Corporation, Kobe MI R&D Center F 6-7-3, Minatojima, Minamimachi, Chuo-ku, Kobe, Hyogo 650-0047, Japan.
  • Nakatani M; Kaneka Corporation, Kobe MI R&D Center F 6-7-3, Minatojima, Minamimachi, Chuo-ku, Kobe, Hyogo 650-0047, Japan.
  • Yoshimura K; Department of Plastic Surgery, School of Medicine, Jichi Medical University, 3311-1 Yakushiji, Shimotsuke, Tochigi 329-0498, Japan.
J Biol Methods ; 3(4): e58, 2016.
Article en En | MEDLINE | ID: mdl-31453220
Fat grafts are valuable for soft-tissue regeneration and augmentation. However, fat graft systems require further improvement for the prediction of graft retention. The concentration of adipose-derived stromal/stem cells (ASCs) is one of the most important factors that affect graft retention; however, current cell quantification techniques have not been applied to adipose tissue. Here we developed a method for the selective quantification of ASCs in tissue (SQAT). We identified a characteristic methylated site in the CD31 promoter after searching for specific markers of ASCs. This DNA methylation was not detected in any cell type other than ASCs in adipose tissue. Therefore, analyzing this methylation may be a suitable approach for quantifying ASCs in tissues because DNA is readily extracted from tissues. SQAT is based on quantifying this methylation by quantitative polymerase chain reaction using methylation-sensitive HapII-treated DNA as the template. SQAT was validated based on the numbers of ASCs determined by CD31-/CD34+-based flow cytometry. The results obtained by both methods were perfectly correlated, thereby demonstrating that SQAT is a useful tool for quantifying ASCs. SQAT analysis using ASCs isolated from suctioned fat according to the standard protocol (i.e., collagenase treatment) showed that the yield of ASCs was 59% ± 21%, which suggests that the ASC isolation technique requires further improvement. Furthermore, SQAT is an excellent method for quantifying ASCs in arbitrary samples (particularly tissue), which could dramatically improve ASC isolation technologies and fat graft systems, thereby facilitating the prediction of graft retention.
Palabras clave
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Tipo de estudio: Guideline / Prognostic_studies Idioma: En Revista: J Biol Methods Año: 2016 Tipo del documento: Article País de afiliación: Japón Pais de publicación: Estados Unidos
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Tipo de estudio: Guideline / Prognostic_studies Idioma: En Revista: J Biol Methods Año: 2016 Tipo del documento: Article País de afiliación: Japón Pais de publicación: Estados Unidos