Choi Y S, Im M W, Kim C S, Lee M H, Noh S E, Lim S M, Kim S L, Cho C G, Kim D I
Department of Biological Engineering, Inha University, Incheon, Korea.
Cytotherapy. 2008;10(2):165-73. doi: 10.1080/14653240701817002.
For successful stem cell-based therapy, not only are alternative good cell sources needed but appropriate scaffolds are key factors. The main purpose of this study was to evaluate the multipotentiality of multilineage progenitor cells (MLPC) and assess the three-dimensional cultivation and chondrogenic differentiation of MLPC in atelocollagen gel for application of tissue-engineered cartilage constructs.
MLPC, human umbilical cord blood-derived clonal cell lines, from BioE Inc. were used. Immunophenotypes of MLPC were characterized using a fluorescence-activated cell sorter (FACS). In vitro differentiation potentials into osteogenic, adipogenic and chondrogenic lineages were examined. Differentiated cells were characterized by reverse transcriptase-polymerase chain reaction (RT-PCR), histologic and immunofluorescence staining. RESULTS Clonogenic MLPC maintained immunophenotypes with specific surface markers of mesenchymal stromal cells (MSC). The osteogenic and adipogenic potentials of MLPC were demonstrated by quantitative real-time PCR, alkaline phosphates activity and Oil Red O staining. Furthermore, MLPC were successfully differentiated into chondrocytes in atelocollagen gel, which was confirmed by RT-PCR and immunofluorescence staining for type II collagen protein.
Whenever MSC are considered for the treatment of cartilage defects, a variety of scaffolds have been utilized as successful carriers for cell delivery. Our results suggest that MLPC can serve as an alternative source for stem cell-based therapy and transplantation. The chondrogenic potential of MLPC in atelocollagen could be suitable for cartilage tissue engineering.
为了实现基于干细胞的成功治疗，不仅需要替代的优质细胞来源，合适的支架也是关键因素。本研究的主要目的是评估多谱系祖细胞（MLPC）的多能性，并评估MLPC在去端胶原蛋白凝胶中的三维培养及软骨分化，以应用于组织工程软骨构建体。
使用来自BioE公司的人脐带血来源的克隆细胞系MLPC。使用荧光激活细胞分选仪（FACS）对MLPC的免疫表型进行表征。检测其向成骨、成脂和软骨谱系的体外分化潜能。通过逆转录聚合酶链反应（RT-PCR）、组织学和免疫荧光染色对分化细胞进行表征。结果 克隆性MLPC维持了间充质基质细胞（MSC）特异性表面标志物的免疫表型。通过定量实时PCR、碱性磷酸酶活性和油红O染色证明了MLPC的成骨和成脂潜能。此外，MLPC在去端胶原蛋白凝胶中成功分化为软骨细胞，这通过RT-PCR和II型胶原蛋白的免疫荧光染色得以证实。
每当考虑将MSC用于治疗软骨缺损时，各种支架已被用作细胞递送的成功载体。我们的结果表明，MLPC可以作为基于干细胞的治疗和移植的替代来源。MLPC在去端胶原蛋白中的软骨生成潜能可能适用于软骨组织工程。
