Isolation of therapeutically functional mouse bone marrow mesenchymal stem cells within 3 h by an effective single-step plastic-adherent method.
Cell Prolif
; 43(3): 235-48, 2010 Jun.
Article
em En
| MEDLINE
| ID: mdl-20546242
ABSTRACT
OBJECTIVES:
Isolation of mouse mesenchymal stem cells (mMSCs), by the approach of plastic adherence, has been difficult due to persistent contamination by haematopoietic cells (HCs); we have observed that this contamination was due to engagement between HCs and mMSCs. The HCs can be lifted together with the mMSCs despite their insensitivity to trypsin digestion. Herein, we provide a single-step procedure to rapidly segregate mMSCs from HC contaminants using transient lower-density plastic adherence (tLDA). MATERIALS ANDMETHODS:
The tLDA was performed by replating bone marrow adherent cells at lower density (1.25 x 10(4) cells/cm(2)) than usual, allowing for transient adherence of no more than 3 h, followed by trypsin digestion. tLDA-isolated cells were evaluated by immunophenotyping, multi-differentiation potentials, immunosuppressive properties, and therapeutic potential as demonstrated by symptoms of osteoporosis.RESULTS:
The single-step tLDA method can effectively eliminate the persistent HC contaminants; tLDA-isolated cells were phenotypically equivalent to those reported as mMSCs. The isolated cells possessed classic tri-lineage differentiation potential into osteogenic, adipogenic and chondrogenic lineages and had immunosuppressive properties. After intravenous transplantation, they migrated into the allogeneic bone marrow and rescued hosts from osteoporosis symptoms, demonstrating their therapeutic potential.CONCLUSIONS:
We have developed a simple and economical method that effectively isolates HC-free, therapeutically functional mMSCs from bone marrow cell adherent cultures. These cells are suitable for various mechanistic and therapeutic studies in the mouse model.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Separação Celular
/
Transplante de Medula Óssea
/
Transplante de Células-Tronco Mesenquimais
/
Células-Tronco Mesenquimais
Limite:
Animals
Idioma:
En
Revista:
Cell Prolif
Ano de publicação:
2010
Tipo de documento:
Article
País de afiliação:
Taiwan