建立人髌下脂肪垫干细胞分离、培养和鉴定的平台，验证其可作为可靠的干细胞来源，并可作为细胞模型参与科学研究。

人髌下脂肪垫取自膝关节置换手术患者，经Ⅰ型胶原酶消化，在高糖Dulbecco改良Eagle培养基(DMEM)内扩增，取P5代细胞用于检测其"干性"及三向分化能力。与微尺度支架结合，通过RT－PCR检测成骨基因的表达，应用最小显著性差异法进行组间两两比较，用于验证微尺度支架的成骨诱导特性。

人髌下脂肪垫干细胞具备间充质干细胞相关特性，包括黏附贴壁性；成脂肪、成软骨及成骨分化；表达相关表面标记，如分化抗原簇(CD)73、CD90和CD105；不表达造血细胞系的分子表面标记，如c-kit、CD14、CD11b、CD34、CD45、CD19、CD79和人白细胞抗原(HLA)-DR。本课题中人髌下脂肪垫来源干细胞成功作为一种干细胞模型，证明HaCG微尺度支架具备成骨诱导特性，表现为三组中成骨基因ALP、COL1、Runx2表达差异有统计学意义(分别为F＝150，P<0.01；F＝68，P<0.01；F＝16，P<0.01)。HaCG微尺度支架组高于纯明胶微尺度支架组及二维培养组，纯明胶微尺度支架组高于二维培养组(P值均小于0.05)。

人髌下脂肪垫干细胞分离、培养和鉴定平台的建立，未来可作为细胞模型应用于多种不同实验。

To establish the platform for isolation, proliferation, identification and application of Human infrapatellar fat pad derived stem cells (IPFP), and to make sure it could be a reliable stem cell model for scientific research.

IPFPs were obtained from patients who underwent knee arthroplasty, then were digested by collagenase I, and expended in high-glucose Dulbecco’s modified Eagle medium (DMEM). The passage five stem cells were used for identifying the stemness and tri-directional differentiation ability. The cells were pipetted on different microscaffolds, to testify the osteoinductivity and biocompatibility of nanohydroxyapatite-chitosan-gelatin microscaffolds (HaCGM) according to the gene expression by Reverse transcription-polymerase chain reaction (RT-PCR). The

were applied least-significant difference (LSD) t-test to perform statistical analysis among groups. Results IPFP-ASCs have the characteristic of mesenchymal stem cells: adherence to plastic; specific surface antigen (Ag) expression, such as cluster of differentiation 73 (CD73), CD90 and CD105; multipotent differentiation potential. HaCGM showed osteogenesis inductivity. The gene expression of ALP, COL1 and Runx2 in three groups were statistically significant(F=150, P < 0.01; F=68, P<0.01; F=16, P<0.01, respectively). HaCGM groups were higher than gelatin groups and 2D culture groups, and gelatin groups were higher than 2D culture groups (P<0.01).

The establishment of the platform for isolation, proliferation, identification and application of Human infrapatellar fat pad derived stem cells, which could be used in different kind of experiments.