Background： Endothelial and smooth muscle cells were used as seeding cells and heterogeneous acellularized matrix was used as scaffold to construct the tissue-engineered graft. Methods： A 2 weeks piglet was selected...Background： Endothelial and smooth muscle cells were used as seeding cells and heterogeneous acellularized matrix was used as scaffold to construct the tissue-engineered graft. Methods： A 2 weeks piglet was selected as a donor of seeding cells. Two-centimetre length of common carotid artery was dissected. Endothelial cells and smooth muscle cells were harvested by trypsin and collagenase digestion respectively. The isolated cells were cultured and expanded using routine cell culture technique. An adult sheep was used as a donor of acellularized matrix. The thoracic aorta was harvested and processed by a multi-step decellularizing technique to remove the original cells and preserve the elastic and collagen fibers. The cultured smooth muscle cells and endothelial cells were then seeded to the acellularized matrix and incubated in vitro for another 2 weeks. The cell seeded graft was then transplanted to the cell-donated piglet to substitute part of the native pulmonary artery. Results： The cultured cells from piglet were characterized as endothelial cells by the presence of specific antigens vWF and CD31, and smooth muscle cells by the presence of specific antigen a-actin on the cell surface respectively with immunohistochemical technique. After decellularizing processing for the thoracic aorta from sheep, all the cellular components were extracted and elastic and collagen fibers kept their original morphology and structure. The maximal load of acellular matrix was decreased and 20% lower than that of untreated thoracic aorta, but the maximal tensions between them were not different statistically and they had similar load-tension curves. Three months after transplantation, the animal was sacrificed and the graft was removed for observation. The results showed that the inner surfaces of the graft were smooth, without thrombosis and calcification. Under microscopy, a great number of growing cells could be seen and elastic and collagen fibers were abundant. Conclusion： Cultured self-derived endothelial and smooth muscle cells could be used as seeding cells and heterogeneous acellularized matrix could be used as scaffold in constructing tissue-engineered graft.展开更多
Objective： To observe the survival and the number of olfactory ensheathing cells （OECs） transplanted in the contused spinal cord, so as to provide a basis for further studying the biological action of OECs.Methods...Objective： To observe the survival and the number of olfactory ensheathing cells （OECs） transplanted in the contused spinal cord, so as to provide a basis for further studying the biological action of OECs.Methods： The rat spinal cords were contused with NYU-impactor Ⅱ at T10 level by dropping a 10 g rod from a height of 25 mm. At the 1st week after injury, OECs isolated freshly from green fluorecense protein （GFP） of the rats were transplanted into the spinal cord at injured site and other two sites 1 mm apart from the caudal and rostral ends with the OECs number of 30000/μl×3 =90000. The survival and the number of OECs were qualitatively and semi-quantitatively observed under the fluorescense microscope from 1 week to 13 weeks after transplantation. The motor function of the cord was evaluated with BBB score.Results： GFP-OECs could survive at least for 13 weeks within the contused spinal cord. Their arrangement was from tight to loose and their number was decreased from 1 week to 13 weeks after injury. The average number of GFP-OECs was 536 at the 1st week, which was less than 1% of the number as compared with original transplantation. After then, the number of GFP-OECs was continually decreased,but the most obvious decrease was found during 1 week to 2 weeks. The extent of decrease at other time points was relatively mild. In contrast to the cell number, motor function of the cord was gradually recovered after transplantation.Conclusions： The survival and the number of GFPOECs are different between the animals and are affected by the pathological reaction of the host cord. Also it is related to the motor function recovery of the contused cord.展开更多
Background:Hepatocellular carcinoma(HCC)is frequently associated withmetabolism dysfunction.Increasing evidence has demonstrated the crucial role of lipidmetabolismin HCC progression.The function of apolipoprotein F(A...Background:Hepatocellular carcinoma(HCC)is frequently associated withmetabolism dysfunction.Increasing evidence has demonstrated the crucial role of lipidmetabolismin HCC progression.The function of apolipoprotein F(ApoF),a lipid transfer inhibitor protein,in HCC is incompletely understood.We aimed to evaluate the functional role of ApoF in HCC in this study.Methods:We used quantitative reverse-transcription polymerase chain reaction(qRT-PCR)to detect ApoF mRNA expression in HCC tissues and hepatoma cell lines(SMMC-7721,HepG2,and Huh7).Immunohistochemistry was performed to detect the expression of ApoF in HCC tissues.The associations between ApoF expression and clinicopathological features as well as HCC prognosis were analyzed.The effect of ApoF on cellular proliferation and growth of SMMC-7721 and Huh7 cells was examined in vitro and in vivo.Results:ApoF expression was significantly down-regulated at both mRNA and protein levels in HCC tissues as compared with adjacent tissues.In SMMC-7721 and Huh7 HCC cells,ApoF overexpression inhibited cell proliferation and migration.In a xenograft nude mouse model,ApoF overexpression effectively controlled HCC growth.Kaplan–Meier analysis results showed that the recurrence-free survival rate of HCC patients with low ApoF expression was significantly lower than that of other HCC patients.Low ApoF expression was associated with several clinicopathological features such as liver cirrhosis,Barcelona Clinic Liver Cancer stage and tumor-node-metastasis stage.Conclusions:ApoF expression was down-regulated in HCC,which was associated with low recurrence-free survival rate.ApoF may serve as a tumor suppressor in HCC and be a potential application for the treatment of this disease.展开更多
基金Project (No. 99ZB14018) supported by the Natural Science Foun-dation of Shanghai, China
文摘Background： Endothelial and smooth muscle cells were used as seeding cells and heterogeneous acellularized matrix was used as scaffold to construct the tissue-engineered graft. Methods： A 2 weeks piglet was selected as a donor of seeding cells. Two-centimetre length of common carotid artery was dissected. Endothelial cells and smooth muscle cells were harvested by trypsin and collagenase digestion respectively. The isolated cells were cultured and expanded using routine cell culture technique. An adult sheep was used as a donor of acellularized matrix. The thoracic aorta was harvested and processed by a multi-step decellularizing technique to remove the original cells and preserve the elastic and collagen fibers. The cultured smooth muscle cells and endothelial cells were then seeded to the acellularized matrix and incubated in vitro for another 2 weeks. The cell seeded graft was then transplanted to the cell-donated piglet to substitute part of the native pulmonary artery. Results： The cultured cells from piglet were characterized as endothelial cells by the presence of specific antigens vWF and CD31, and smooth muscle cells by the presence of specific antigen a-actin on the cell surface respectively with immunohistochemical technique. After decellularizing processing for the thoracic aorta from sheep, all the cellular components were extracted and elastic and collagen fibers kept their original morphology and structure. The maximal load of acellular matrix was decreased and 20% lower than that of untreated thoracic aorta, but the maximal tensions between them were not different statistically and they had similar load-tension curves. Three months after transplantation, the animal was sacrificed and the graft was removed for observation. The results showed that the inner surfaces of the graft were smooth, without thrombosis and calcification. Under microscopy, a great number of growing cells could be seen and elastic and collagen fibers were abundant. Conclusion： Cultured self-derived endothelial and smooth muscle cells could be used as seeding cells and heterogeneous acellularized matrix could be used as scaffold in constructing tissue-engineered graft.
文摘Objective： To observe the survival and the number of olfactory ensheathing cells （OECs） transplanted in the contused spinal cord, so as to provide a basis for further studying the biological action of OECs.Methods： The rat spinal cords were contused with NYU-impactor Ⅱ at T10 level by dropping a 10 g rod from a height of 25 mm. At the 1st week after injury, OECs isolated freshly from green fluorecense protein （GFP） of the rats were transplanted into the spinal cord at injured site and other two sites 1 mm apart from the caudal and rostral ends with the OECs number of 30000/μl×3 =90000. The survival and the number of OECs were qualitatively and semi-quantitatively observed under the fluorescense microscope from 1 week to 13 weeks after transplantation. The motor function of the cord was evaluated with BBB score.Results： GFP-OECs could survive at least for 13 weeks within the contused spinal cord. Their arrangement was from tight to loose and their number was decreased from 1 week to 13 weeks after injury. The average number of GFP-OECs was 536 at the 1st week, which was less than 1% of the number as compared with original transplantation. After then, the number of GFP-OECs was continually decreased,but the most obvious decrease was found during 1 week to 2 weeks. The extent of decrease at other time points was relatively mild. In contrast to the cell number, motor function of the cord was gradually recovered after transplantation.Conclusions： The survival and the number of GFPOECs are different between the animals and are affected by the pathological reaction of the host cord. Also it is related to the motor function recovery of the contused cord.
基金This study was supported by grants from the National Natural Science Foundation of China[No.81572726]the Natural Science Foundation of Guangdong Province[No.2018A030313641 and No.2016A030313848]+1 种基金the Science and Technology Planning Project of Guangdong Province[No.2014A020212122 and No.2016A020212004]the Medical Research Foundation of Guangdong Province[No.A2016312].
文摘Background:Hepatocellular carcinoma(HCC)is frequently associated withmetabolism dysfunction.Increasing evidence has demonstrated the crucial role of lipidmetabolismin HCC progression.The function of apolipoprotein F(ApoF),a lipid transfer inhibitor protein,in HCC is incompletely understood.We aimed to evaluate the functional role of ApoF in HCC in this study.Methods:We used quantitative reverse-transcription polymerase chain reaction(qRT-PCR)to detect ApoF mRNA expression in HCC tissues and hepatoma cell lines(SMMC-7721,HepG2,and Huh7).Immunohistochemistry was performed to detect the expression of ApoF in HCC tissues.The associations between ApoF expression and clinicopathological features as well as HCC prognosis were analyzed.The effect of ApoF on cellular proliferation and growth of SMMC-7721 and Huh7 cells was examined in vitro and in vivo.Results:ApoF expression was significantly down-regulated at both mRNA and protein levels in HCC tissues as compared with adjacent tissues.In SMMC-7721 and Huh7 HCC cells,ApoF overexpression inhibited cell proliferation and migration.In a xenograft nude mouse model,ApoF overexpression effectively controlled HCC growth.Kaplan–Meier analysis results showed that the recurrence-free survival rate of HCC patients with low ApoF expression was significantly lower than that of other HCC patients.Low ApoF expression was associated with several clinicopathological features such as liver cirrhosis,Barcelona Clinic Liver Cancer stage and tumor-node-metastasis stage.Conclusions:ApoF expression was down-regulated in HCC,which was associated with low recurrence-free survival rate.ApoF may serve as a tumor suppressor in HCC and be a potential application for the treatment of this disease.
