Comparative study on seeding methods of human bone marrow stromal cells in bone tissue engineering.
BACKGROUND
In general the traditional static seeding method has its limitation while the dynamic seeding method reveals its advantages over traditional static method. We compared static and dynamic seeding method for human bone marrow stromal cells (hBMSCs) in bone tissue engineering.


METHODS
DNA assay was used for detecting the maximal initial seeding concentration for static seeding. Dynamic and static seeding methods were compared, when scaffolds were loaded with hBMSCs at this maximal initial cell seeding concentration. Histology and scanning electron microscope (SEM) were examined to evaluate the distribution of cells inside the constructs. Markers encoding osteogenic genes were measured by fluorescent RT-PCR. The protocol for dynamic seeding of hBMSCs was also investigated.


RESULTS
DNA assay showed that the static maximal initial seeding concentration was lower than that in dynamic seeding. Histology and SEM showed even distribution and spread of cells in the dynamically seeded constructs, while their statically seeded counterparts showed cell aggregation. Fluorescent RT-PCR again showed stronger osteogenic potential of dynamically seeded constructs.


CONCLUSIONS
dynamic seeding of hBMSCs is a promising technique in bone tissue engineering.
Qi
Xin
X
Department of Orthopedic Surgery, First Hospital of Jilin University, Changchun 130021, China. xinqi@public.cc.il.cn
Liu
Jian-Guo
JG
Chang
Ying
Y
Xu
Xin-Xiang
XX
D001854:Bone Marrow Cells / Q000166:cytology*; D002478:Cells, Cultured; D004247:DNA / Q000032:analysis; D006801:Humans; D008854:Microscopy, Electron; D010012:Osteogenesis; D011108:Polymers; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017154:Stromal Cells / Q000166:cytology; D023822:Tissue Engineering / Q000379:methods*
D011108:Polymers; D004247:DNA