Bioreactor expansion of human adult bone marrow-derived mesenchymal stem cells.
Supplementation of mesenchymal stem cells (MSCs) during hematopoietic stem cell (HSC) transplantation alleviates complications such as graft-versus-host disease, leading to a speedy recovery of hematopoiesis. To meet this clinical demand, a fast MSC expansion method is required. In the present study, we examined the feasibility of using a rotary bioreactor system to expand MSCs from isolated bone marrow mononuclear cells. The cells were cultured in a rotary bioreactor with Myelocult medium containing a combination of supplementary factors, including stem cell factor and interleukin-3 and -6. After 8 days of culture, total cell numbers, Stro-1(+)CD44(+)CD34(-) MSCs, and CD34(+)CD44(+)Stro-1(-) HSCs were increased 9-, 29-, and 8-fold, respectively. Colony-forming efficiency-fibroblast per day of the bioreactor-treated cells was 1.44-fold higher than that of the cells without bioreactor treatment. The bioreactor-expanded MSCs showed expression of primitive MSC markers endoglin (SH2) and vimentin, whereas markers associated with lineage differentiation, including osteocalcin (osteogenesis), type II collagen (chondrogenesis), and C/EBP-alpha (CCAAT/enhancer-binding protein-alpha) (adipogenesis), were not detected. Upon induction, the bioreactor-expanded MSCs were able to differentiate into osteoblasts, chondrocytes, and adipocytes. We conclude that the rotary bioreactor with the modified Myelocult medium reported in this study may be used to rapidly expand MSCs.
Chen
Xi
X
Department of Orthopaedic Surgery, School of Biomedical Sciences, Queen's University Belfast, Musgrave Park Hospital, Belfast, BT9 7JB, United Kingdom.
Xu
Haibo
H
Wan
Chao
C
McCaigue
Mervyn
M
Li
Gang
G
D000368:Aged; D000369:Aged, 80 and over; D019149:Bioreactors; D001854:Bone Marrow Cells / Q000166:cytology*; D002452:Cell Count; D019070:Cell Lineage; D049109:Cell Proliferation; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D003470:Culture Media; D005347:Fibroblasts / Q000166:cytology; D006801:Humans; D007700:Kinetics; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D008875:Middle Aged; D010641:Phenotype; D011506:Proteins / Q000378:metabolism; D013997:Time Factors
D003470:Culture Media; D011506:Proteins