Clinical-scale cultures of cord blood CD34(+) cells to amplify committed progenitors and maintain stem cell activity.
Cell Transplant
; 20(9): 1453-63, 2011.
Article
en En
| MEDLINE
| ID: mdl-21294956
We developed a clinical-scale cord blood (CB) cell ex vivo procedure to enable an extensive expansion of committed progenitors--colony-forming cells (CFCs) without impairing very primitive hematopoietic stem cells (HSCs). CD34(++) cells, selected from previously cryopreserved and thawed CB units, were cultured in two steps (diluted 1:4 after 6 days) in the presence of stem cell factor (SCF), fms-related tyrosine kinase 3 ligand (Flt-3L), megakaryocyte growth and development factor (MGDF) (100 ng/ml each), granulocyte-colony stimulating factor (G-CSF) (10 ng/ml) in HP01 serum-free medium. HSC activity was evaluated in a serial transplantation assay, by detection of human cells (CD45, CD33, CD19 and CFC of human origin) in bone marrow (BM) of primary and secondary recipient NOD/SCID mice 6-8 weeks after transplantation. A wide amplification of total cells (â¼350-fold), CD34(+) cells (â¼100-fold), and CFC (â¼130-fold) without impairing the HSC activity was obtained. The activity of a particular HSC subpopulation (SRC(CFC)) was even enhanced.Thus, an extensive ex vivo expansion of CFCs is feasible without impairing the activity of HSCs. This result was enabled by associating antioxidant power of medium with an appropriate cytokine cocktail (i.e., mimicking physiologic effects of a weak oxygenation in hematopoietic environment).
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Células Madre
/
Linaje de la Célula
/
Técnicas de Cultivo de Célula
/
Antígenos CD34
/
Sangre Fetal
Límite:
Animals
/
Humans
Idioma:
En
Revista:
Cell Transplant
Asunto de la revista:
TRANSPLANTE
Año:
2011
Tipo del documento:
Article
País de afiliación:
Francia
Pais de publicación:
Estados Unidos