Osteogenic potential of human bone marrow-derived mesenchymal stromal cells cultured in autologous serum: a preliminary study.
OBJECTIVE
As part of the authors' research on potential osteogenesis by filling bone defects with human bone marrow-derived mesenchymal stromal cells (hBM-MSCs) in patients with cleft lip and palate, they examined the cytoproliferative potential and cytobiological activity of hBM-MSCs in vitro and their osteogenic potential in vivo without performing osteoinduction.


METHODS
The hBM-MSCs were collected from iliac cancellous bone and then used in primary culture, followed by 2 subcultures using an autologous serum (AS)-containing medium and a fetal bovine serum (FBS)-containing medium. Cytoproliferative potential and cytobiological activity as expressed by bone markers (alkaline phosphatase and osteocalcin) in hBM-MSCs cultured in the AS-containing medium (AS-cultured hBM-MSCs) and the FBS-containing medium (FBS-cultured hBM-MSCs) were examined in vitro, and the osteogenic potential of AS- and FBS-cultured hBM-MSCs was examined in mice.


RESULTS
On day 6 of the second subculture, the number of hBM-MSCs per milliliter of specimen from 8 pediatric patients was significantly larger (P < .05) in FBS-cultured compared with AS-cultured hBM-MSCs. The alkaline phosphatase activity of hBM-MSCs was significantly greater (P < .05) when cultured in the AS-containing medium compared with the FBS-containing medium. The in vivo study showed the formation of an osteoid-like matrix rather than definite bone tissue.


CONCLUSIONS
1) FBS is appropriate for the cytoproliferation of hBM-MSCs; 2) the AS-containing medium is likely to have a good possibility of inducing the differentiation of hBM-MSCs; and 3) AS-cultured hBM-MSCs contain a group of cells that spontaneously differentiate into an osteoid-like matrix without performing osteoinduction.
Department of Plastic and Aesthetic Surgery, School of Medicine, Kitasato University, Kanagawa, Japan.
Akira
A
Takeda
Yasuharu
Y
Yamazaki
Kyoko
K
Baba
Masashi
M
Ishiguro
Kazuya
K
Aoyagi
Shigehiro
S
Ikemoto
Eiju
E
Uchinuma
D000469:Alkaline Phosphatase / Q000032:analysis; D000818:Animals; D015415:Biomarkers / Q000032:analysis; D001769:Blood; D001854:Bone Marrow Cells / Q000145:classification / Q000502:physiology*; D001857:Bone Matrix / Q000166:cytology / Q000502:physiology; D002452:Cell Count; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D002648:Child; D003470:Culture Media; D017886:Durapatite; D005260:Female; D006801:Humans; D008297:Male; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000145:classification / Q000502:physiology*; D051379:Mice; D008819:Mice, Nude; D008928:Mitochondria / Q000145:classification; D015675:Osteocalcin / Q000032:analysis; D010012:Osteogenesis / Q000502:physiology*; D040521:Subcutaneous Tissue / Q000601:surgery; D054457:Tissue Scaffolds
D015415:Biomarkers; D003470:Culture Media; D015675:Osteocalcin; D017886:Durapatite; D000469:Alkaline Phosphatase