| 黄闯宋镜明宋扬刘佳王丽颖金作林△.犬牙囊干细胞膜片的构建及生物学特性的研究[J].,2012,12(2):226-230 |
| 犬牙囊干细胞膜片的构建及生物学特性的研究 |
| Construction of Dental Follicle Stem Cell Sheet and its BiologicalCharacteristics in Beagle Dogs |
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| DOI： |
| 中文关键词: 牙囊干细胞(DFSCs) 细胞膜片 分化 |
| 英文关键词: Dental Follicle Stem Cells(DFSCs) Cell sheet Differentiation |
| 基金项目:国家自然科学基金（30973358） |
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| 中文摘要: |
| 目的：利用犬牙囊干细胞（Dental Follicle Stem Cells, DFSCs）构建细胞膜片并研究其生物学特性。方法：取4 至6 月龄犬尖牙
牙胚，分离培养DFSCs，鉴定。用含抗坏血酸的培养基诱导2 周构建细胞膜片，并通过倒置显微镜、HE 染色、茜素红染色、油红染
色、扫描电镜(SEM)对膜片进行形态学检测，检测成骨、成脂能力。结果：DFSCs 于体外被成功分离、纯化、培养，细胞克隆形成率约
为5.1%。流式鉴定为CD29+CD44+CD34-，增殖能力及克隆形成能力较强，并能成功构建成细胞膜片。光镜和电镜显示膜片细胞
排列紧密，细胞基质分泌多，油红O 染色后可见细胞内有大量脂滴形成。(B) 茜素红染色后可见大量清晰的钙结节形成。结论：成
功构建犬DFSCs 膜片，并证明其具有较强的成骨能力，为进一步利用犬DFSCs 膜片修复牙槽骨缺损的研究提供条件。 |
| 英文摘要: |
| Objective: To study the construction of dental follicle stem cell sheet and its biological characteristics in Beagle dogs.
Methods: After identification,DFSCs were sub-cultured to construct DFSCs sheet. Cell sheet was investaged by inverted microscope, HE
staining and scanning electron microscope (SEM). DFSCs sheets were induced by adipogenesis inducing medium and osteogenic medium
for 14 days separately. Oil red staining and Alizarin red staining was applied to examine adipogenic induction and osteogenic induction.
Results: DFSCs showed typical spindle shape.. Colony-forming assay results showed about 5.1% DFSCs colony formation. DFSCs were
positive for CD29 and CD44, but negative for CD34. MTT manifested the growth and proliferation was good. Cell cycle testing showed:
G1=87.1%,G2=5.54%. DFSCs sheets were constructed successfully and its growth in multilayer. It found that DFSCs expanded adequately
and extracellular matrix(ECM) was clear and numerous in scanning electron micrescopy.Oil red staining and alizarin red staining
both demonstrated positive reactions in DFSCs sheet after induction. Conclusion: It suggested that DFSCs cell sheet may be constructed
and has a strong bone-forming ability. |
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