Though Yurman province contains some 562 known species of fish, no cell lines from any of these have been made available to date. To protect germplasm resources and provide an effective tool in solving problems at cel...Though Yurman province contains some 562 known species of fish, no cell lines from any of these have been made available to date. To protect germplasm resources and provide an effective tool in solving problems at cellular level of Anabarilius grahami, a fish endemic to Fuxian Lake, Yunnan, China, we established and characterized the major features of a continuous cell line （AGF II） from the caudal fin tissue of A. grahami. This AGF II cell line consists of fibroblast-like cells and has been subeultured more than 60 times over the course of a year. The cell line was maintained in DMEM/F12 supplemented with 10% FBS, with a cellular doubling time of 51.1 h. We continued with more experiments to optimize the culture and storage conditions, and found a variety of interesting results： cells could grow at temperature between 24 ~C and 28 ~C, with the optimal temperature of 28 ~C. Likewise, the growth rate ofA. grahami fin cells increased when the FBS proportion increased from 5% to 20%, with the optimal growth at the concentrations of 20% FBS, cells were able to grow in L-15 and DMEM/FI2 with optimal growth at L-15; DMSO is a better eryoprotectant than Glycerol, EG and MeOH for AGFII cells with optimal concentration of 5% DMSO. Chromosome analysis also showed that the distribution of chromosome number varies from 38 to 52, with a modal peak at 48 chromosomes, accounting for 39.8% of all cells. Using the same primer pairs specific to mtDNA, the AGF II cell sequences obtained by PCR were identical to those from muscle tissues ofA. grahami. Both chromosome analysis and PCR amplification confirmed the AGF II cells were from A. grahami, also indicating that that current long-term artificial propagation ofA. grahami has been successful. Finally, we noted that when cells were transfected with pEYFP-N1 and pECFP-N1 plasmid, bright fluorescent signals were observed, suggesting that this cell line may be suitable for use in transfection and future gene expression studies.展开更多
AIM： To evaluate the presence of progenitor cells in healthy adult rat liver displaying the equivalent ad- vanced hepatogenic profile as that obtained in humans. METHODS： Rat fibroblastic-like liver derived cells （...AIM： To evaluate the presence of progenitor cells in healthy adult rat liver displaying the equivalent ad- vanced hepatogenic profile as that obtained in humans. METHODS： Rat fibroblastic-like liver derived cells （rFLDC） were obtained from collagenase-isolated liver cell suspensions and characterized and their phenotype profile determined using flow cytometry, immunocyto- chemistry, reverse transcription polymerase chain reac- tion and functional assays. RESULTS： rFLDC exhibit fibroblastoid morphology, ex- press mesenchymal （CD73, CD90, vimentin, m-smooth muscle actin）, hepatocyte （UGTIA1, CK8） and biliary （CK19） markers. Moreover, these cells are able to store glycogen, and have glucose 6 phosphatase activity, but not UGTIA1 activity. Under the hepatogenic differentia- tion protocol, rFLDC display an up-regulation of hepatocyte markers expression （albumin, tryptophan 2,3-di- oxygenase, G6Pase） correlated to a down-regulation of the expression of the biliary marker CK19. CONCLUSION： Advanced hepatic features observed in human liver progenitor cells could not be demonstrated in rFLDC. However, we demonstrated the presence of an original rodent hepato-biliary cell type.展开更多
Objective： To study the preparative method of controlled release microspheres incorporating basic fibroblast growth factor （bFGF） and the bioaetivities of bFGF, which were released from bFGF mierospheres, on the cu...Objective： To study the preparative method of controlled release microspheres incorporating basic fibroblast growth factor （bFGF） and the bioaetivities of bFGF, which were released from bFGF mierospheres, on the cultured Sehwann cells. Methods： bFGF was microcapsulated with the multiple emulsion encapsulative method using polylactic-coglycolic acid （PLGA） as coating material. Its morphology, particle size distribution, drug loading, enveloping rate and in vitro release property were studied. The cultured Schwann cells were grouped according to the different ingredients being added to the culture medium of bFGF group or bFGF-PLGA group. Then the cytometry, cytoactivity detection and mitotic cycle analysis of Schwann cells were performed. Results： The morphology and the particle size distribution of the bFGF-PLGA microspheres were even and good. The drug loading and enveloping rate of microspheres were （ 27.18×10^-3 ） % ± （0.51×10^-3） % and 66.43 % ± 1.24 %. The release property of microspheres in vitro was good and the overall release rate was 72. 47 % in 11 days. The in vitro cellular study showed that： at the first 2 days of plate culture, the cell number and viability of the bFGF group were statistically higher than the bFGF-PLGA group; at the 3rd and 4th days of plate culture, the cell number and viability of bFGF and bFGF-PLGA groups showed no difference; at the 6th and 8th days of the plate culture, the cell number and viability of the bFGF-PLGA group were statistically higher than the bFGF group. By flow eytometry examination, at the 2nd day of plate culture, the G2/M ＋ S percentage of bFGF group was statistically higher than the bFGF-PLGA group, at the 4th and 8th days of plate culture, the G2/M ＋ S percentage of the bFGF-PLGA group was statistically higher than the bFGF group. Conclusions： It is practical to prepare the bFGF- PLGA microspheres with the multiple emulsion eneapsulative method, bFGF-PLGA mierospheres can preserve the bioaetivities of bFGF effectively and promote the proliferation of Sehwann cells in a long period because of the controlled release of bFGF from the mierospheres.展开更多
The tumor microenvironment （TME） plays an important role in supporting cancer progression. The TME is composed of tumor cells, the surrounding tumor-associated stromal cells, and the extracellular matrix （ECM）. Cr...The tumor microenvironment （TME） plays an important role in supporting cancer progression. The TME is composed of tumor cells, the surrounding tumor-associated stromal cells, and the extracellular matrix （ECM）. Crosstalk between the TME components contributes to tumorigenesis. Recently, one of our studies showed that pancreatic ductal adenocarcinoma （PDAC） cells can induce DNA methylation in cancer-associated flbroblasts （CAFs）, thereby modifying tumor-stromal interactions in the TME, and subsequently creating a TME that supports tumor growth Here we summarize recent studies about how DNA methylation affects tumorigenesis through regulating tumor- associated stromal components including fibroblasts and immune cells. We also discuss the potential for targeting DNA methylation for the treatment of cancers.展开更多
文摘Though Yurman province contains some 562 known species of fish, no cell lines from any of these have been made available to date. To protect germplasm resources and provide an effective tool in solving problems at cellular level of Anabarilius grahami, a fish endemic to Fuxian Lake, Yunnan, China, we established and characterized the major features of a continuous cell line （AGF II） from the caudal fin tissue of A. grahami. This AGF II cell line consists of fibroblast-like cells and has been subeultured more than 60 times over the course of a year. The cell line was maintained in DMEM/F12 supplemented with 10% FBS, with a cellular doubling time of 51.1 h. We continued with more experiments to optimize the culture and storage conditions, and found a variety of interesting results： cells could grow at temperature between 24 ~C and 28 ~C, with the optimal temperature of 28 ~C. Likewise, the growth rate ofA. grahami fin cells increased when the FBS proportion increased from 5% to 20%, with the optimal growth at the concentrations of 20% FBS, cells were able to grow in L-15 and DMEM/FI2 with optimal growth at L-15; DMSO is a better eryoprotectant than Glycerol, EG and MeOH for AGFII cells with optimal concentration of 5% DMSO. Chromosome analysis also showed that the distribution of chromosome number varies from 38 to 52, with a modal peak at 48 chromosomes, accounting for 39.8% of all cells. Using the same primer pairs specific to mtDNA, the AGF II cell sequences obtained by PCR were identical to those from muscle tissues ofA. grahami. Both chromosome analysis and PCR amplification confirmed the AGF II cells were from A. grahami, also indicating that that current long-term artificial propagation ofA. grahami has been successful. Finally, we noted that when cells were transfected with pEYFP-N1 and pECFP-N1 plasmid, bright fluorescent signals were observed, suggesting that this cell line may be suitable for use in transfection and future gene expression studies.
基金Supported by Fonds pour la formation à la recherche dans l’industrie et dans l’agriculture (FRIA)
文摘AIM： To evaluate the presence of progenitor cells in healthy adult rat liver displaying the equivalent ad- vanced hepatogenic profile as that obtained in humans. METHODS： Rat fibroblastic-like liver derived cells （rFLDC） were obtained from collagenase-isolated liver cell suspensions and characterized and their phenotype profile determined using flow cytometry, immunocyto- chemistry, reverse transcription polymerase chain reac- tion and functional assays. RESULTS： rFLDC exhibit fibroblastoid morphology, ex- press mesenchymal （CD73, CD90, vimentin, m-smooth muscle actin）, hepatocyte （UGTIA1, CK8） and biliary （CK19） markers. Moreover, these cells are able to store glycogen, and have glucose 6 phosphatase activity, but not UGTIA1 activity. Under the hepatogenic differentia- tion protocol, rFLDC display an up-regulation of hepatocyte markers expression （albumin, tryptophan 2,3-di- oxygenase, G6Pase） correlated to a down-regulation of the expression of the biliary marker CK19. CONCLUSION： Advanced hepatic features observed in human liver progenitor cells could not be demonstrated in rFLDC. However, we demonstrated the presence of an original rodent hepato-biliary cell type.
基金This study was supported by the National Natural Science Foundation of China （No. 39970747）.
文摘Objective： To study the preparative method of controlled release microspheres incorporating basic fibroblast growth factor （bFGF） and the bioaetivities of bFGF, which were released from bFGF mierospheres, on the cultured Sehwann cells. Methods： bFGF was microcapsulated with the multiple emulsion encapsulative method using polylactic-coglycolic acid （PLGA） as coating material. Its morphology, particle size distribution, drug loading, enveloping rate and in vitro release property were studied. The cultured Schwann cells were grouped according to the different ingredients being added to the culture medium of bFGF group or bFGF-PLGA group. Then the cytometry, cytoactivity detection and mitotic cycle analysis of Schwann cells were performed. Results： The morphology and the particle size distribution of the bFGF-PLGA microspheres were even and good. The drug loading and enveloping rate of microspheres were （ 27.18×10^-3 ） % ± （0.51×10^-3） % and 66.43 % ± 1.24 %. The release property of microspheres in vitro was good and the overall release rate was 72. 47 % in 11 days. The in vitro cellular study showed that： at the first 2 days of plate culture, the cell number and viability of the bFGF group were statistically higher than the bFGF-PLGA group; at the 3rd and 4th days of plate culture, the cell number and viability of bFGF and bFGF-PLGA groups showed no difference; at the 6th and 8th days of the plate culture, the cell number and viability of the bFGF-PLGA group were statistically higher than the bFGF group. By flow eytometry examination, at the 2nd day of plate culture, the G2/M ＋ S percentage of bFGF group was statistically higher than the bFGF-PLGA group, at the 4th and 8th days of plate culture, the G2/M ＋ S percentage of the bFGF-PLGA group was statistically higher than the bFGF group. Conclusions： It is practical to prepare the bFGF- PLGA microspheres with the multiple emulsion eneapsulative method, bFGF-PLGA mierospheres can preserve the bioaetivities of bFGF effectively and promote the proliferation of Sehwann cells in a long period because of the controlled release of bFGF from the mierospheres.
基金supported by the National Cancer Institute(Nos.R01CA169702,P50CA062924,and R01CA197296),the United States
文摘The tumor microenvironment （TME） plays an important role in supporting cancer progression. The TME is composed of tumor cells, the surrounding tumor-associated stromal cells, and the extracellular matrix （ECM）. Crosstalk between the TME components contributes to tumorigenesis. Recently, one of our studies showed that pancreatic ductal adenocarcinoma （PDAC） cells can induce DNA methylation in cancer-associated flbroblasts （CAFs）, thereby modifying tumor-stromal interactions in the TME, and subsequently creating a TME that supports tumor growth Here we summarize recent studies about how DNA methylation affects tumorigenesis through regulating tumor- associated stromal components including fibroblasts and immune cells. We also discuss the potential for targeting DNA methylation for the treatment of cancers.
