文档介绍：华中科技大学
博士学位论文
碳纳米纤维支架与兔脂肪干细胞的体外相容性研究
姓名:萨伊德
申请学位级别:博士
专业:外科学(泌尿外科)
指导教师:叶章群;曾晓勇
2011-04
华中科技大学博士学位论文

patibility of Carbon Nanofibrous Scaffold and Rabbit
Adipose-Derived Stem Cells in Vitro
Abstract
Part I Characteristics of Isolation and Culture of Rabbit
Adipose-Derived Stem Cells in vitro.
Objectives: To establish the method for isolation and culture of rabbit adipose-derived
stem cells (ADSCs) in vitro, identifying & analyzing its biological characteristics, and
exploring the optimal condition for culture of adipose-derived stem cells in vitro.
Methods: Three female New Zealand rabbits were selected, the adipose-derived stem cells
in inguinal region were harvested by digestion of collagenase I enzymatic hydrolysis. After
the differential adherent culture conducted to the selected passage cells, the cells extracted
and purified. The flow cytometry is adopted to indentify adipose-derived stem cells using
monoclonal antibodies of mouse-anti-rabbit CD44 and CD166. The proliferation and
growth curve of the 4th generation of ADSCs were detected to investigate the
morphological features and proliferation states of the primary and passage adipose-derived
stem cells (ADSCs), while adipogenic and osteogenic inductions and identification were
also implemented. ADSCs were induced for differentiation in osteogenic medium for 3 w
and in adipogenic medium for 2 w. Finally, the target genes were detected once a week
using RT-PCR technique.
Results:
The growth morphology of primary and passaged ADSCs : the morphology of primary
cell was smaller and grows adherently in the shape of long-spindle or polygon; the shape of
passage cell achieved by differential adherent culture was expanded in cobble-stone-like
distribution, and the growth rate of the passage cell that is in active growth and migration
was found significantly faster than primary cell.
The result of flow cytometry shows that the positive rate of