组织工程与重建外科杂志
組織工程與重建外科雜誌
조직공정여중건외과잡지
JOURNAL OF TISSUE ENGINEERING AND RECONSTRUCTIVE SURGERY
2013年
4期
181-185
,共5页
刘李娜%何爱娟%周广东%曹卫刚
劉李娜%何愛娟%週廣東%曹衛剛
류리나%하애연%주엄동%조위강
骨髓间充质干细胞%胎猪%组织工程%软骨%种子细胞
骨髓間充質榦細胞%胎豬%組織工程%軟骨%種子細胞
골수간충질간세포%태저%조직공정%연골%충자세포
Bone marrow mesenchymal stem cells%Porcine fetal%Tissue engineering%Cartilage%Seed cell
目的比较胎猪骨髓间充质干细胞（Bone Marrow Mesenchymal Stem Cells，BMSCs）和成年猪BMSCs构建软骨能力的差异，寻找合适的同种异体组织工程软骨种子细胞来源。方法通过剖腹产手术获得胎龄为70 d的胎猪，胎猪骨髓液贴壁培养获得胎猪BMSCs；抽取成年猪骨髓液，经贴壁培养法获得成年猪BMSCs。两种细胞体外扩增培养后,观察第3代细胞形态，并进行成骨、成脂和成软骨诱导。分别取两种细胞以1×108 cells/mL的细胞终浓度，接种于聚乳酸包埋的聚羟基乙酸支架，体外诱导培养8周后取材。通过大体观察、糖胺聚糖（GAG）含量测定、总胶原含量测定、组织学，以及免疫组化等方法，对两种细胞构建的组织工程软骨的相关生物学特性进行比较。结果胎猪BMSCs比成年猪BMSCs具有更好的增殖和成骨、成脂和成软骨能力。胎猪BMSCs构建的软骨有良好的软骨外观，而且GAG含量和总胶原含量均高于成年猪BMSCs构建的软骨（P<0.01）。组织学和免疫组化显示，胎猪BMSCs构建的软骨组织结构致密，基质及Ⅱ型胶原显色程度均明显强于成年猪BMSCs构建的软骨。结论胎猪BMSCs是组织工程软骨较好的种子细胞来源。
目的比較胎豬骨髓間充質榦細胞（Bone Marrow Mesenchymal Stem Cells，BMSCs）和成年豬BMSCs構建軟骨能力的差異，尋找閤適的同種異體組織工程軟骨種子細胞來源。方法通過剖腹產手術穫得胎齡為70 d的胎豬，胎豬骨髓液貼壁培養穫得胎豬BMSCs；抽取成年豬骨髓液，經貼壁培養法穫得成年豬BMSCs。兩種細胞體外擴增培養後,觀察第3代細胞形態，併進行成骨、成脂和成軟骨誘導。分彆取兩種細胞以1×108 cells/mL的細胞終濃度，接種于聚乳痠包埋的聚羥基乙痠支架，體外誘導培養8週後取材。通過大體觀察、糖胺聚糖（GAG）含量測定、總膠原含量測定、組織學，以及免疫組化等方法，對兩種細胞構建的組織工程軟骨的相關生物學特性進行比較。結果胎豬BMSCs比成年豬BMSCs具有更好的增殖和成骨、成脂和成軟骨能力。胎豬BMSCs構建的軟骨有良好的軟骨外觀，而且GAG含量和總膠原含量均高于成年豬BMSCs構建的軟骨（P<0.01）。組織學和免疫組化顯示，胎豬BMSCs構建的軟骨組織結構緻密，基質及Ⅱ型膠原顯色程度均明顯彊于成年豬BMSCs構建的軟骨。結論胎豬BMSCs是組織工程軟骨較好的種子細胞來源。
목적비교태저골수간충질간세포（Bone Marrow Mesenchymal Stem Cells，BMSCs）화성년저BMSCs구건연골능력적차이，심조합괄적동충이체조직공정연골충자세포래원。방법통과부복산수술획득태령위70 d적태저，태저골수액첩벽배양획득태저BMSCs；추취성년저골수액，경첩벽배양법획득성년저BMSCs。량충세포체외확증배양후,관찰제3대세포형태，병진행성골、성지화성연골유도。분별취량충세포이1×108 cells/mL적세포종농도，접충우취유산포매적취간기을산지가，체외유도배양8주후취재。통과대체관찰、당알취당（GAG）함량측정、총효원함량측정、조직학，이급면역조화등방법，대량충세포구건적조직공정연골적상관생물학특성진행비교。결과태저BMSCs비성년저BMSCs구유경호적증식화성골、성지화성연골능력。태저BMSCs구건적연골유량호적연골외관，이차GAG함량화총효원함량균고우성년저BMSCs구건적연골（P<0.01）。조직학화면역조화현시，태저BMSCs구건적연골조직결구치밀，기질급Ⅱ형효원현색정도균명현강우성년저BMSCs구건적연골。결론태저BMSCs시조직공정연골교호적충자세포래원。
Objective To investigate the optimal seed cell for cartilage engineering by comparing the chondrogenesis capability of porcine fetal bone marrow mesenchymal stem cells (fBMSCs) and adult porcine BMSCs (aBMSCs). Methods Fetal pigs with gestational age of 70 days were obtained by the uterine-incision delivery, and primary fBMSCs were isolated from the bone marrow. Primary aBMSCs were isolated from the bone marrow which was aspirated from iliac. Cell morphology of the two kinds of cells at passage 3 were observed after in vitro proliferation. The two kinds of cells at passages 3 were characterized by their osteogenic, adipogenic and chondrogenic differentiation potential. Then the fBMSCs and aBMSCs were separately seeded onto a polyglycolic acid/polylactic acid (PGA/PLA) scaffold with the concentration of 1 ×108 cells/mL. All specimens were harvested after 8 weeks' culture in vitro. Gross observation, glycosaminoglycan (GAG) quantification, total collagen quantification and histology were used to compare related characteristic differences of engineered cartilage formed by the two kinds of cells. Results fBMSCs had better proliferation and multiple differentiation capacity than aBMSCs. The two kinds of cells both formed mature cartilage after 8 weeks of culture in vitro, and the engineered cartilage of aBMSCs group had better appearance. The GAG content and total collagen content of the cartilage formed by fBMSCs were both higher than the cartilage formed by aBMSCs (P<0.01). Histology and immunohistochemistry demonstrated that the cartilage formed by fBMSCs have more compact tissue structure. The cartilage matrix staining of cartilage formed by fBMSCs were stronger than that of cartilage formed by aBMSCs. Conclusion The fBMSCs seems to be the optimal seed cells for cartilage tissue engineering.
