摘要
分别采用动态种植(旋转烧瓶法)和静态种植方法种植人骨髓间质干细胞,于种植后的两周内测定细胞-载体构件中DNA含量;利用组织学光镜和扫描电镜观察细胞的分布情况;运用荧光标记RT-PCR技术测定相关成骨基因的表达。构件中DNA含量测定表明,对于静态种植,当初始种植密度为每载体400×103(8.9×104/mm3)时,DNA的含量达到最高;在此基础上提高初始种植密度,并不能进一步提高构件DNA含量。光镜和扫描电镜观察可见动态种植后人骨髓间质干细胞在载体中的分布相对均匀,静态种植后细胞在载体中出现聚集现象;荧光标记RT-PCR证明,体外构件培养两周后,动态种植后的细胞-载体构件中有较多的成骨基因表达。提示人骨髓间质干细胞的静态种植效率较低;动态种植是一种优于静态种植的可行方法。
Seeding is the crucial step in bone tissue engineering. In current study, static and dynamic seeding methods for human bone marrow stem cells （hBMSCs） were compared. The methods for assay of DNA content in the constructs after seeding were adopted. The optic microscopy for histological apearance and the scanning electron microscopy（SEM） and fluorescent RT-PCR for osteogenic markers were performed, The maximal initial seeding concentration in static seeding is lower than that in dynamic seeding, Histology and SEM revealed the even distribution and spreading of cells in the dynamically seeded constructs, but showed cell aggregation in the statically seeded counterparts, Fluorescent RT-PCR again revealed stronger osteogenic potential of dynamically seeded constructs. Therefore, this initial study demonstrated that dynamic seeding of human bone marrow stem cells is a promising technique in bone tissue engineering.
出处
《生物医学工程学杂志》
EI
CAS
CSCD
北大核心
2006年第1期175-179,共5页
Journal of Biomedical Engineering
关键词
组织工程
骨再生
人骨髓间质干细胞
动态种植
Tissue engineering Bone regeneration Human bone marrow stem cell （hBMSC）Dynamic seeding
