Aim To characterize the odontogenic capability of apical bud and phenotypical change of apical bud cells （ABCs） in different microenvironment.Methodology Incisor apical bud tissues from neonatal SD rat were dissecte...Aim To characterize the odontogenic capability of apical bud and phenotypical change of apical bud cells （ABCs） in different microenvironment.Methodology Incisor apical bud tissues from neonatal SD rat were dissected and transplanted into the renal capsules to determine their odontogenic capability. Meanwhile ABCs were cultured and purified by repeated differential trypsinization. Then ABCs were cultured with conditioned medium from developing apical complex cells （DAC-CM）. Immunocytochemistry, reverse transcriptase polymerase chain reaction （RT-PCR） and scanning electron microscope （SEM） were performed to compare the biolo- gical change of ABC treated with or without DAC-CM. Results First we confirmed the ability of apical bud to form crown-like structure ectopically. Equally important, by using the developing apical complex （DAC） condi- tioned medium, we found the microenvironment created by root could abrogate the ＂crown＂ features of ABCs and promote their proliferation and differentiation. Conclusion ABCs possess odontogenic capability to form crown-like tissues and this property can be affected by root-produced microenvironment.展开更多
目的:建立一种分离、培养大鼠切牙apical bud上皮细胞的有效方法。方法:选择出生后5 d SD仔鼠,分离下颌切牙牙胚。将切牙颈部末端apical bud切下,酶消化后原代培养。经差别消化及选择性培养液纯化,并对培养的细胞进行角蛋白14和釉原蛋...目的:建立一种分离、培养大鼠切牙apical bud上皮细胞的有效方法。方法:选择出生后5 d SD仔鼠,分离下颌切牙牙胚。将切牙颈部末端apical bud切下,酶消化后原代培养。经差别消化及选择性培养液纯化,并对培养的细胞进行角蛋白14和釉原蛋白、波形丝蛋白免疫组织化学染色鉴定。结果:培养的原代细胞混杂部分间充质细胞,经差别消化和选择性培养液纯化后获得了单一的上皮样细胞,此类细胞角蛋白14和釉原蛋白染色阳性,波形丝蛋白染色阴性。结论:经本方法的分离、培养成功地获得了纯化的大鼠切牙颈环上皮细胞。展开更多
基金supported by National Nature Science Foundation of China(Project No.3057 2046,30725042)
文摘Aim To characterize the odontogenic capability of apical bud and phenotypical change of apical bud cells （ABCs） in different microenvironment.Methodology Incisor apical bud tissues from neonatal SD rat were dissected and transplanted into the renal capsules to determine their odontogenic capability. Meanwhile ABCs were cultured and purified by repeated differential trypsinization. Then ABCs were cultured with conditioned medium from developing apical complex cells （DAC-CM）. Immunocytochemistry, reverse transcriptase polymerase chain reaction （RT-PCR） and scanning electron microscope （SEM） were performed to compare the biolo- gical change of ABC treated with or without DAC-CM. Results First we confirmed the ability of apical bud to form crown-like structure ectopically. Equally important, by using the developing apical complex （DAC） condi- tioned medium, we found the microenvironment created by root could abrogate the ＂crown＂ features of ABCs and promote their proliferation and differentiation. Conclusion ABCs possess odontogenic capability to form crown-like tissues and this property can be affected by root-produced microenvironment.
文摘目的:建立一种分离、培养大鼠切牙apical bud上皮细胞的有效方法。方法:选择出生后5 d SD仔鼠,分离下颌切牙牙胚。将切牙颈部末端apical bud切下,酶消化后原代培养。经差别消化及选择性培养液纯化,并对培养的细胞进行角蛋白14和釉原蛋白、波形丝蛋白免疫组织化学染色鉴定。结果:培养的原代细胞混杂部分间充质细胞,经差别消化和选择性培养液纯化后获得了单一的上皮样细胞,此类细胞角蛋白14和釉原蛋白染色阳性,波形丝蛋白染色阴性。结论:经本方法的分离、培养成功地获得了纯化的大鼠切牙颈环上皮细胞。
