Observation and quantitative analysis of rat bone marrow stromal cells cultured in vitro on newly formed transparent beta-tricalcium phosphate.
To observe living cell morphology on ceramics by light microscopy, we fabricated a new material-transparent beta - tricalcium phosphate (t-beta TCP) ceramic-for the purpose of serving as a tissue culture substrate. Bone marrow stromal cells (BMSCs) were obtained from rat femora and cultured on both t-beta TCP ceramic disks and culture grade polystyrene (PS) dishes in an osteogenic medium. After 1 day of culture, cell attachment and spreading on both the t-beta TCP and PS substrata were equally and clearly detected by ordinary light microscopy. After 14 days of culture, extensive cell growth, alkaline phosphatase (ALP) staining, and bone mineral deposition could be detected on both substrata. In addition, quantitative biochemical analyses revealed high DNA content, ALP activity, and osteocalcin content of these cultures. This experiment is significant in that all of the results were similarly observed on both the t-beta TCP and PS substrata, indicating the excellent properties of beta TCP ceramics for BMSCs culture towards osteogenic differentiation.
Kotobuki
Noriko
N
Research Institute for Cell Engineering (RICE), National Institute of Advanced Industrial Science and Technology (AIST), 3-11-46 Nakouji, Amagasaki, Hyogo, 661-0974, Japan. n.kotobuki@aist.go.jp
Kawagoe
Daisuke
D
Nomura
Daishirou
D
Katou
Youichi
Y
Muraki
Kaori
K
Fujimori
Hirotaka
H
Goto
Seishi
S
Ioku
Koji
K
Ohgushi
Hajime
H
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D001672:Biocompatible Materials / Q000378:metabolism*; D001854:Bone Marrow Cells / Q000166:cytology*; D002130:Calcium Phosphates / Q000378:metabolism*; D002452:Cell Count; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D004247:DNA / Q000032:analysis; D005269:Femur / Q000166:cytology; D008297:Male; D015675:Osteocalcin / Q000378:metabolism; D011137:Polystyrenes / Q000737:chemistry; D051381:Rats; D011916:Rats, Inbred F344; D017154:Stromal Cells / Q000166:cytology* / Q000378:metabolism*
D001672:Biocompatible Materials; D002130:Calcium Phosphates; D011137:Polystyrenes; C485817:beta-tricalcium phosphate; D015675:Osteocalcin; D004247:DNA; D000469:Alkaline Phosphatase