CAJ | 학술논문

背景:实验证实鹿茸多肽可以促进体外培养软骨细胞的增殖和细胞外基质糖胺多糖、Ⅱ型胶原、Aggrecan蛋白的表达.目的:通过对体外培养的兔骨髓间充质干细胞在特定培养液作用下向软骨细胞表型分化的研究,探讨鹿茸多肽对其软骨分化的影响.方法:将第3代兔骨髓间充质干细胞随机分为空白对照组、诱导组、鹿茸多肽组,分别采用普通培养液、诱导培养液、含10 mg/L鹿茸多肽的诱导培养液于离心管内进行培养;并取兔的关节软骨细胞作为关节软骨组.分别于1,2,3周后取材,通过组织学、生物化学和RT-PCR技术,对离心管内构建的软骨组织进行形态学和细胞功能状态的观察.结果与结论:空白对照组培养2周后,细胞团块逐渐崩解,无法进行苏木精-伊红染色.诱导组、鹿茸多肽组细胞团块除有轻度收缩外,呈白色半透明状;苏木精-伊红染色发现部分细胞为圆形或卵圆形,表层细胞密度大;诱导组、鹿茸多肽组糖胺多糖含量及Ⅱ型胶原mRNA表达随培养时间延长而增多,各时间点诱导组、鹿茸多肽组含量均高于空白对照组(P < 0.05);各时间点鹿茸多肽组糖胺多糖含量及Ⅱ型胶原mRNA表达均高于诱导组,但低于关节软骨组 (P < 0.05).提示骨髓间充质干细胞在特定培养条件下能向软骨细胞表型分化,且鹿茸多肽对其定向软骨分化有明显促进作用.虽然在体外可以构建出软骨组织,但其与关节软骨质量相比仍有很大差距.
배경:실험증실록용다태가이촉진체외배양연골세포적증식화세포외기질당알다당、Ⅱ형효원、Aggrecan단백적표체.목적:통과대체외배양적토골수간충질간세포재특정배양액작용하향연골세포표형분화적연구,탐토록용다태대기연골분화적영향.방법:장제3대토골수간충질간세포수궤분위공백대조조、유도조、록용다태조,분별채용보통배양액、유도배양액、함10 mg/L록용다태적유도배양액우리심관내진행배양;병취토적관절연골세포작위관절연골조.분별우1,2,3주후취재,통과조직학、생물화학화RT-PCR기술,대리심관내구건적연골조직진행형태학화세포공능상태적관찰.결과여결론:공백대조조배양2주후,세포단괴축점붕해,무법진행소목정-이홍염색.유도조、록용다태조세포단괴제유경도수축외,정백색반투명상;소목정-이홍염색발현부분세포위원형혹란원형,표층세포밀도대;유도조、록용다태조당알다당함량급Ⅱ형효원mRNA표체수배양시간연장이증다,각시간점유도조、록용다태조함량균고우공백대조조(P < 0.05);각시간점록용다태조당알다당함량급Ⅱ형효원mRNA표체균고우유도조,단저우관절연골조 (P < 0.05).제시골수간충질간세포재특정배양조건하능향연골세포표형분화,차록용다태대기정향연골분화유명현촉진작용.수연재체외가이구건출연골조직,단기여관절연골질량상비잉유흔대차거.
BACKGROUND: Pilose antler polypeptides (PAP) have been proved to promote the proliferation of condrocytes cultured in vitro and expressions of glycosaminoglycan (GAG), type Ⅱ collagen and Aggrecan protein in the extracellular matrix.OBJECTIVE: To investigate the feasibility of chondrogenic phenotype differentiation of rabbit bone marrow-derived mensenchymal stem cells (BMSCs) in a defined medium and then to study the effect of PAP on chondrogenic differentiation of BMSCs in vitro.METHODS: The third passage BMSCs from rabbits were randomly divided into control group cultured in ordinary medium, induced group cultured in defined medium, and PAP group cultured in defined medium containing 10 mg/L PAP. An equal volume of articular chondrocytes were selected from rabbits as articular cartilage group. The cellular morphological and functional characteristics were observed after 1, 2, 3 weeks in centrifuge tubes by histological, biochemical and reverse transcription-polymerase chain reaction (RT-PCR) technique. RESULTS AND CONCLUSION: Cell masses in the control group gradually crumbled after 2 weeks, and hematoxylin-eosin staining could not be done. Cell masses in the induced and PAP groups were semitransparent, but slightly contracted. A part of these cells were round or oval with a high density distribution at the surface. The content of GAG and mRNA expression of type Ⅱ collagen in the induced and PAP groups were increased with culture time, and higher than those in the control group at different time points (P < 0.05). The content of GAG and mRNA expression of type Ⅱ collagen in the PAP group were higher than those in the induced group, but lower than those in the articular cartilage group (P < 0.05). The results indicated that BMSCs can differentiate into chondrogenic phenotype in the defined medium, and PAP can significantly enhance chondrogenic phenotype differentiation of BMSCs. But the quality of cultured cartilage tissue is poorer than that of the articular cartilage.