CAJ | 학술논문

背景:随着新生生血清的产业化,新生生跟腱资源大量积累,使得利用其制备医用胶原蛋白海绵并产业化成为可能.目的:采用新生生跟腱制备胶原蛋白海绵,观察其与Veto细胞、天祝白牦生胚胎皮肤细胞和岷县黑紫羔羊睾丸细胞的生物相容性.设计、时间、地点:重复测量观察,于2006-02105在西北民族人学生命科学与工程学院生物工程与技术国家民委重点实验室完成.材料:用出生24 h内的新生生跟腱经冰醋酸、胃蛋白酶等消化.经盐析、透析及冻干等处理后,制成胶原蛋白海绵.天祝白牦生胚胎皮肤f2代细胞和岷县黑紫羔羊睾丸f2代细胞为自制,Veto细胞为协和医科人学细胞中心提供.方法:在6孔板中,将3种细胞分别接种于经紫外线、臭氧灭菌后的胶原蛋白海绵上,于37℃、体积分数为0.05的CO2恒温箱培养,同时设对照实验.主要观察指标:相差显微镜观察接种5,12,24,72 h时3种细胞在胶原蛋白海绵上和6孔板底的生K情况,接种11 d对共培养物行考马斯亮蓝,苏术精一伊红染色.结果:接种后5 h.在胶原蛋白海绵周围的6孔板底部可看到细胞已贴壁、伸展,个别细胞有分裂现象,在胶原蛋白海绵表面,隐约可见圆彤细胞排列,接种后48～72 h时,6孔板底部细胞铺满单层.接种后11 d,考马斯亮蓝和苏小精一伊红染色显示3种共培养物中均有大量细胞良好生长,胶原海绵外观变得挺拔、透明、有韧性.结论:新生生跟腱胶原蛋向海绵与上述3种来自不同动物、不同组织的细胞有很好的生物相容性,可做皮肤细胞、肾细胞和睾丸细胞的培养支架.
배경:수착신생생혈청적산업화,신생생근건자원대량적루,사득이용기제비의용효원단백해면병산업화성위가능.목적:채용신생생근건제비효원단백해면,관찰기여Veto세포、천축백모생배태피부세포화민현흑자고양고환세포적생물상용성.설계、시간、지점:중복측량관찰,우2006-02105재서북민족인학생명과학여공정학원생물공정여기술국가민위중점실험실완성.재료:용출생24 h내적신생생근건경빙작산、위단백매등소화.경염석、투석급동간등처리후,제성효원단백해면.천축백모생배태피부f2대세포화민현흑자고양고환f2대세포위자제,Veto세포위협화의과인학세포중심제공.방법:재6공판중,장3충세포분별접충우경자외선、취양멸균후적효원단백해면상,우37℃、체적분수위0.05적CO2항온상배양,동시설대조실험.주요관찰지표:상차현미경관찰접충5,12,24,72 h시3충세포재효원단백해면상화6공판저적생K정황,접충11 d대공배양물행고마사량람,소술정일이홍염색.결과:접충후5 h.재효원단백해면주위적6공판저부가간도세포이첩벽、신전,개별세포유분렬현상,재효원단백해면표면,은약가견원동세포배렬,접충후48～72 h시,6공판저부세포포만단층.접충후11 d,고마사량람화소소정일이홍염색현시3충공배양물중균유대량세포량호생장,효원해면외관변득정발、투명、유인성.결론:신생생근건효원단향해면여상술3충래자불동동물、불동조직적세포유흔호적생물상용성,가주피부세포、신세포화고환세포적배양지가.
BACKGROUND:Industrialization of new-born bovine serum and abundant resource of bovine lendon enable industrialization of medical collagen sponge.OBJECTIVE:To prepare collagen sponge with new-born bovine tendons by inoculating Veto cells,primary embryo skin cell of Tianzhu White Yak and lamb testicle cell of Minxian black fur sheep of the tissue scaffold of collagen sponge.and observe the biocompatibility of collagen sponge with three different cells.DESIGN,TIME AND SETTING:Repetitive measurement was performed at the Key Laboratory of State Nationalities Afrairs Committee,College of Life Sciences and Engineering,Northwest University for Nationalities from February to May 2006.MATERIALS:Tendons of new-born bovine within 24 hours were digested by glacial acetic acid and pepsinum firstly and then salting-out,dialysis and vacum freeze drying were performed to prepare collagen sponge.f2 passage of embryo skin cells of Tianzhu Whit Yak and f2 passage of lamb testicle cells of Minxian black fur sheep were prepared by out laboratory;Vero cells were provided by Union Medical University.METHODS:In a 6-hole plate,the Vero cell,embryo skin cells of Tianzhu White Yak and lamb testicle cells of Minxian Black Fur Sheep were inoculated into the collagen sponge pretreated by ultraviolet and sterilized by ozone.and incubated in 5%CO2 at 37℃.In addition,cells only inoculated ia a culture plate served as control. MAIN OUTCOME MEASURES:Inverted phase contrast microscope was used to observe cell growth condition in the collagen sponge and 6-hole plate at 5,12,24 and 72 hours.In addition,Coomassie brilliant blue as well as HE staining were conducted at 11 days after culture to identify the culture.RESULTS:Five hours after inoculation,cell adherence and expansion was observed at the bottom of culture plate.and some of the cells showed division.On the surface of collagen sponge.a round cell arrangement was observed.After inoculation of 48-72 hours,monolayer was found at the bottom of the plate.On the 11th day of culture.Coomassie brilliant blue and HE staining of three kinds of cells showed there were lager amount of cells well grew in the holes of collagen sponge,and the collagen sponge turned to be eminent,transparent and tenacious.CONCLUSION:The collagen sponge made from new-born bovine tendons exhibit good biocompatibility with three kinds of cells from different animals and tissues,and can be served as culture seaffoId of skin cells,tenal ceils.and testicle cells.