关键词: 脂肪组织, 基质细胞, 细胞培养, 成肌分化

Abstract:

Objective　To isolate human adipose tissue-derived stromal cells and study the potential of osteogenic differentiation after inductive culture.Methods　Liposuction human adipose tissueswere minced and digested with collagenase typeⅠ. The ob- tained stromal cellswere plated in BGJbmedium as primary culture forten days. The second passage cellswere harvested and plat- ed in DMEM/F12 medium supplemented with 10% FBS, 5% horse serum and 50μmol/L hydrocortisone for myogenic induction culture. The cell-anchored slips were removed and fixed in 4% formaldehydum polymerisatum. Toluidine blue, Mallory′s phos- photungstic hematoxylin staining and monoclonal antibody to human skeletal muscle myosin heavy chain immunocytochemical meth- ods were used to assay the differentiation of cells.Results　It was observed that the size and shape of induced cells were much different from those of non-induced cells. Toluidine blue, Mallory′s phosphotungstic hematoxylin staining demonstrated there were many basophilic striations within cytoplasm and multinucleated myotubes were formed. Immunocytochemical stain indicated that characterastic skeletal myosin heavy chain was positive in myogenic induced cells.Conclusion　It seems that human adipose tis- sue represents an abundant reservoir of adult stem cells that have multi-germline potential to differentiate into myoblasts. Adipose tissue derived stromal cells will be another alternative source for cell-based tissue engineering in skeletal muscle reconstruction.

Key words: adipose tissue, stromal cell, cell culture, myogenic differentiation