Mesenchymal stem cells(MSCs)have received significant attention in recent years due to their large potential for cell therapy.Indeed,they secrete a wide variety of immunomodulatory factors of interest for the treatmen...Mesenchymal stem cells(MSCs)have received significant attention in recent years due to their large potential for cell therapy.Indeed,they secrete a wide variety of immunomodulatory factors of interest for the treatment of immune-related disorders and inflammatory diseases.MSCs can be extracted from multiple tissues of the human body.However,several factors may restrict their use for clinical applications:the requirement of invasive procedures for their isolation,their limited numbers,and their heterogeneity according to the tissue of origin or donor.In addition,MSCs often present early signs of replicative senescence limiting their expansion in vitro,and their therapeutic capacity in vivo.Due to the clinical potential of MSCs,a considerable number of methods to differentiate induced pluripotent stem cells(iPSCs)into MSCs have emerged.iPSCs represent a new reliable,unlimited source to generate MSCs(MSCs derived from iPSC,iMSCs)from homogeneous and well-characterized cell lines,which would relieve many of the above mentioned technical and biological limitations.Additionally,the use of iPSCs prevents some of the ethical concerns surrounding the use of human embryonic stem cells.In this review,we analyze the main current protocols used to differentiate human iPSCs into MSCs,which we classify into five different categories:MSC Switch,Embryoid Body Formation,Specific Differentiation,Pathway Inhibitor,and Platelet Lysate.We also evaluate common and method-specific culture components and provide a list of positive and negative markers for MSC characterization.Further guidance on material requirements to produce iMSCs with these methods and on the phenotypic features of the iMSCs obtained is added.The information may help researchers identify protocol options to design and/or refine standardized procedures for large-scale production of iMSCs fitting clinical demands.展开更多
Objective:At present, human umbilical cord mesenchymal stem cells (hucMSC) have been widely used in basic research and clinical trials in treatment of many diseases, but how to obtain a large number and high quality o...Objective:At present, human umbilical cord mesenchymal stem cells (hucMSC) have been widely used in basic research and clinical trials in treatment of many diseases, but how to obtain a large number and high quality of stable and reliable mesenchymal stem cells for clinical application is still a scientific research needs to be resolved. Since each laboratory used different culture methods, the productivity of the different cultures may vary. Therefore, the purpose of this study is to optimize a stable and reliable hucMSC culture procedure. Methods: Mesenchymal stem cells derived from human umbilical cord were isolated and cultured in different culture medium. By comparing the morphology, passage, proliferation and using flow cytometry to detect the immunophenotypes of the cells, optimized cultural method was selected.Results: By comparison of the time of cells migrating out from the tissue, cell growth status, cell number and expression of CD90 andCD73 of the hucMSCs, we found that X-VIVO15+10% fetal bovine serum group, X-VIVO15+2% fetal bovine serum and 2% UltroserG group, and MSC basal medium+UltraGRO-Advanced group had an earlier migration time from the umbilical cord and shorter confluence time to cover 80%-90% of the cuturing flask. The Huc-MSCs cultured with MSC basal medium+2% UltraGRO-Advanced group revealed the fastest proliferation. The expression of CD90 and CD73 in Huc-MSCs cultured with MSC basal medium+2% UltraGRO-Advanced group were above 90%, while the expression of CD90 and CD73 in other groups were below 90%. The expression of CD34 and CD45 surface antigens were low in huc-MSCs from all the groups.Conclusion:In summary, MSC basal medium-UltraGRO-Advanced group was the optimal culture medium for human umbilical cord mesenchymal stem cells.展开更多
To establish and validate a method for cell irradiation in 96-well and 6-well plates using a linear accelerator, three irradiation methods(G0 B0 F40,G0 B1.5 F40, and G180 B1.5 F40) were designed to irradiate cell cult...To establish and validate a method for cell irradiation in 96-well and 6-well plates using a linear accelerator, three irradiation methods(G0 B0 F40,G0 B1.5 F40, and G180 B1.5 F40) were designed to irradiate cell culture plasticware simulated with RW3 slab phantom and polystyrene. The difference between the actual physical measured dose and the preset dose was compared among the three methods under the preparatory conditions of 2, 4, 6, 8, and 10 Gy. MDA-MB-231 cells were analyzed by using a cell proliferation assay and a clonogenic assay to verify the difference between the three cell irradiation methods on cell radiosensitivity. For each preset dose, the difference between the actual measured dose and the preset dose was the lowest for Method G0 B1.5 F40, the second lowest for Method G180 B1.5 F40, and the maximum for Method GOB0 F40. The ranges of the differences were-0.28 to 0.02%,-2.17 to-1.80%, and-4.92 to-4.55%, and 0.31 to-0.12%,-3.42 to-2.86%, and-7.31 to-6.92%,respectively, for 96-well and 6-well plates. The cell culture experiments proved that Method G0 B1.5 F40 was an accurate, effective, simple, and practical irradiation method. The most accurate and effective cell irradiation method should always be used, as it will reduce dose differences and instability factors and provide improved accuracy and comparability for laboratories researching cellular radiosensitivity.展开更多
AIM: Activated pancreatic stellate cells (PSCs) have been implicated in the pathogenesis of pancreatic fibrosis and inflammation. Primary PSCs can be subcultured only several times because of their limited growth pote...AIM: Activated pancreatic stellate cells (PSCs) have been implicated in the pathogenesis of pancreatic fibrosis and inflammation. Primary PSCs can be subcultured only several times because of their limited growth potential. A continuous cell line may therefore be valuable in studying molecular mechanisms of these pancreatic disorders. The aim of this study was to establish a cell line of rat PSCs by spontaneous immortalization.METHODS: PSCs were isolated from the pancreas of male Wistar rats, and conventional subcultivation was performed repeatedly. Telomerase activity was measured using the telomere repeat amplification protocol. Activation of transcription factors was assessed by electrophoretic mobility shift assay.Activation of mitogen-activated protein (MAP) kinases was examined by Western blotting using anti-phosphospecific antibodies. Expression of cytokine-induced neutrophil chemoattractant-1 was determined by enzyme immunoassay.RESULTS: Conventional subcultivation yielded actively growing cells. One clone was obtained after limiting dilution,and designated as SIPS. This cell line has been passaged repeatedly more than 2 years, and is thus likely immortalized.SIPS cells retained morphological characteristics of primary,culture-activated PSCs. SIPS expressed α-smooth muscle actin, glial acidic fibrillary protein, vimentin, desmin, type Ⅰ collagen, fibronectin, and prolyl hydroxylases. Telomerase activity and p53 expression were negative. Proliferation of SIPS cells was serum-dependent, and stimulated with platelet-derived growth factor-BB through the activation of extracellular signal-regulated kinase. Interleukin-1β activated nuclear factor-κB, activator protein-1, and MAP kinases.Interleukin-1β induced cytokine-induced neutrophil chemoattractant-1 expression through the activation of nuclear factor-κB and MAP kinases.CONCLUSION: SIPS cells can be useful for in vitro studies of cell biology and signal transduction of PSCs.展开更多
INTRODUCTIONCrypt epithelial cells in normal small intestineproliferate at a high speed. But they are verydifficult to culture in vitro and passage stably. A lotof studies have been done[1-16]. Some domestic labsisola...INTRODUCTIONCrypt epithelial cells in normal small intestineproliferate at a high speed. But they are verydifficult to culture in vitro and passage stably. A lotof studies have been done[1-16]. Some domestic labsisolated and cultured crypt cells from embryonalintestines and aseptic animal intestine, but failed.We introduced normal rat epithelial cell line-IEC-6from the USA and its living condition for stablepassage was successfully established after trials. Thecell line was testified to be the small intestinalepithelial cell by electron microscopy,immunihistochemistry and enzymatic histoch-emistry. It has been applied to some relatedresearch work[17-21]. It was found that manyfactors were involved in the culture system. Ourpresent study focuses on the culture method and theinfluencing factors on IEC-6.展开更多
Preliminary animal experiments have confirmed that sensory nerve fibers promote osteoblast differentiation, but motor nerve fibers have no promotion effect. Whether sensory neurons pro- mote the proliferation and oste...Preliminary animal experiments have confirmed that sensory nerve fibers promote osteoblast differentiation, but motor nerve fibers have no promotion effect. Whether sensory neurons pro- mote the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells remains unclear. No results at the cellular level have been reported. In this study, dorsal root ganglion neurons （sensory neurons） from Sprague-Dawley fetal rats were co-cultured with bone marrow mesenchymal stem cells transfected with green fluorescent protein 3 weeks after osteo- genic differentiation in vitro, while osteoblasts derived from bone marrow mesenchymal stem cells served as the control group. The rat dorsal root ganglion neurons promoted the prolifera- tion of bone marrow mesenchymal stem cell-derived osteoblasts at B and 5 days of co-culture, as observed by fluorescence microscopy. The levels of mRNAs for osteogenic differentiation-re- lated factors （including alkaline phosphatase, osteocalcin, osteopontin and bone morphogenetic protein 2） in the co-culture group were higher than those in the control group, as detected by real-time quantitative PCR. Our findings indicate that dorsal root ganglion neurons promote the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells, which pro- vides a theoretical basis for in vitro experiments aimed at constructing tissue-engineered bone.展开更多
Objective\ To test the effect of hypothermia on Na+/H+ exchange, activated by shrinkage and cytoplasmic acidosis. Method\ Amiloride-sensitive Na+ influx in guinea pig red cells was traced with isotope 22Na an...Objective\ To test the effect of hypothermia on Na+/H+ exchange, activated by shrinkage and cytoplasmic acidosis. Method\ Amiloride-sensitive Na+ influx in guinea pig red cells was traced with isotope 22Na and intracellular Na+ concentration was measured by emission flame photometry. Result\ Amiloride-sensitive Na+ influx decreased linearly as a function of temperatures (about 37℃) in shrunken cells, but increased in acidified cells. The up-regulation of acid-induced Na+/H+ exchange by elevated temperature was enhanced by hypo-osmolarity. Less sensitivity of intracellular H+ site at 41℃ may be the mechanism for the inhibition of shrinkage-induced Na+/H+ exchange by elevated temperature. Heating-mediated explosive increase in the activity of acid-induced Na+/H+ exchange may be due to enhanced extracellular Na+ sensitivity and lower intracellular pH caused by acidic metabolites. Acid-induced Na+/H+ ewxchange contributes to cytoplasmic Na+ accumulation. Conclusion\ These two modes of Na+/H+ exchange with different response to elevated temperature may play different roles in the cellular pathogenesis of heatstroke.展开更多
To assess, in vitro, the cellular viability in a murine macrophage cell line J774 with 9 different orthodontic wires and to evaluate the effects of its NO production. To assess cellular viability by MTT: 3-(4,5-dimeth...To assess, in vitro, the cellular viability in a murine macrophage cell line J774 with 9 different orthodontic wires and to evaluate the effects of its NO production. To assess cellular viability by MTT: 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl tetrazolium bromide assay in the cell line J774 with 9 different orthodontic wires and quantify NO production by these macrophages. Cell cultures were evaluated at 24, 48 and 72 hours. There was no significant difference of the means of cellular viability between the control and the group of wires in the respective time intervals. In the comparison with the control group, there was significant difference in the NO production in groups 1, 6, and 9 at 24 hours interval. Group 8 showed significant difference in relation to the control group at final time interval. Cellular viability in all groups was higher at the final time interval than at the initial time interval. This increase was significant in the control group. In the material groups, the final mean of cellular viability at 72 hours showed no significant difference when compared with the control group. NO production in all groups was higher at the final time interval than at the initial time interval. This increase was significant in the control group. In the material groups, the final mean of NO production at 72 hours was only significant in group 8 (betatitanium) when compared with the control group.展开更多
The human dermis presents an ongoing problem for regenerative medicine. Current medical management uses various acellular dermal matrices on wound sites. The challenge for scientists is to examine, then to question ac...The human dermis presents an ongoing problem for regenerative medicine. Current medical management uses various acellular dermal matrices on wound sites. The challenge for scientists is to examine, then to question accepted conventional wisdom and to present new concepts. In this paper, Autologous Cell Therapy will be described by using cell culture of autologous dermal fibroblasts and their extracellular matrix as a foundation for rebuilding the dermis in conditioned wound beds. This proposal seems to create a conflict with the medical approach to keeping a wound bed “moist”.展开更多
OBJECTIVE: To investigate the proliferation and differentiation of cultured corneal stem cells and determine the effect of corneal stem cells cultured on amniotic membranes on the limbal area for treating corneal burn...OBJECTIVE: To investigate the proliferation and differentiation of cultured corneal stem cells and determine the effect of corneal stem cells cultured on amniotic membranes on the limbal area for treating corneal burns. METHODS: The proliferation and differentiation of corneal stem cells in vitro had been examined using colony-forming efficiency and immunohistochemistry. The stem cells had been cultured on amniotic membranes and transplanted to the limbal area for treating corneal burns. RESULTS: Corneal stem cells had a high proliferation capacity in primary and first passage, cytokeratin 3 was not expressed in primary culture but partly in first passage. The stem cells could proliferate to form cell layer on an amniotic membrane. When transplanted, stem cells could survive on limbus. After transplantation, ocular inflammation resolved, the cornea re-epithelialized, the stromal opacity reduced, the superficial neovascularity was lessened and the conjunctival fornix re-established. CONCLUSIONS: Ocular surface conditions could be improved by allograft of corneal stem cells cultured on amniotic membranes.展开更多
文摘Mesenchymal stem cells(MSCs)have received significant attention in recent years due to their large potential for cell therapy.Indeed,they secrete a wide variety of immunomodulatory factors of interest for the treatment of immune-related disorders and inflammatory diseases.MSCs can be extracted from multiple tissues of the human body.However,several factors may restrict their use for clinical applications:the requirement of invasive procedures for their isolation,their limited numbers,and their heterogeneity according to the tissue of origin or donor.In addition,MSCs often present early signs of replicative senescence limiting their expansion in vitro,and their therapeutic capacity in vivo.Due to the clinical potential of MSCs,a considerable number of methods to differentiate induced pluripotent stem cells(iPSCs)into MSCs have emerged.iPSCs represent a new reliable,unlimited source to generate MSCs(MSCs derived from iPSC,iMSCs)from homogeneous and well-characterized cell lines,which would relieve many of the above mentioned technical and biological limitations.Additionally,the use of iPSCs prevents some of the ethical concerns surrounding the use of human embryonic stem cells.In this review,we analyze the main current protocols used to differentiate human iPSCs into MSCs,which we classify into five different categories:MSC Switch,Embryoid Body Formation,Specific Differentiation,Pathway Inhibitor,and Platelet Lysate.We also evaluate common and method-specific culture components and provide a list of positive and negative markers for MSC characterization.Further guidance on material requirements to produce iMSCs with these methods and on the phenotypic features of the iMSCs obtained is added.The information may help researchers identify protocol options to design and/or refine standardized procedures for large-scale production of iMSCs fitting clinical demands.
基金National Natural Science Foundation(:81570157)Shandong Province Medical and Health Technology Development Project(2013ws0291).
文摘Objective:At present, human umbilical cord mesenchymal stem cells (hucMSC) have been widely used in basic research and clinical trials in treatment of many diseases, but how to obtain a large number and high quality of stable and reliable mesenchymal stem cells for clinical application is still a scientific research needs to be resolved. Since each laboratory used different culture methods, the productivity of the different cultures may vary. Therefore, the purpose of this study is to optimize a stable and reliable hucMSC culture procedure. Methods: Mesenchymal stem cells derived from human umbilical cord were isolated and cultured in different culture medium. By comparing the morphology, passage, proliferation and using flow cytometry to detect the immunophenotypes of the cells, optimized cultural method was selected.Results: By comparison of the time of cells migrating out from the tissue, cell growth status, cell number and expression of CD90 andCD73 of the hucMSCs, we found that X-VIVO15+10% fetal bovine serum group, X-VIVO15+2% fetal bovine serum and 2% UltroserG group, and MSC basal medium+UltraGRO-Advanced group had an earlier migration time from the umbilical cord and shorter confluence time to cover 80%-90% of the cuturing flask. The Huc-MSCs cultured with MSC basal medium+2% UltraGRO-Advanced group revealed the fastest proliferation. The expression of CD90 and CD73 in Huc-MSCs cultured with MSC basal medium+2% UltraGRO-Advanced group were above 90%, while the expression of CD90 and CD73 in other groups were below 90%. The expression of CD34 and CD45 surface antigens were low in huc-MSCs from all the groups.Conclusion:In summary, MSC basal medium-UltraGRO-Advanced group was the optimal culture medium for human umbilical cord mesenchymal stem cells.
基金supported by the Hospital Personnel Climbing Plan of the Tenth People's Hospital Affiliated to Tongji University
文摘To establish and validate a method for cell irradiation in 96-well and 6-well plates using a linear accelerator, three irradiation methods(G0 B0 F40,G0 B1.5 F40, and G180 B1.5 F40) were designed to irradiate cell culture plasticware simulated with RW3 slab phantom and polystyrene. The difference between the actual physical measured dose and the preset dose was compared among the three methods under the preparatory conditions of 2, 4, 6, 8, and 10 Gy. MDA-MB-231 cells were analyzed by using a cell proliferation assay and a clonogenic assay to verify the difference between the three cell irradiation methods on cell radiosensitivity. For each preset dose, the difference between the actual measured dose and the preset dose was the lowest for Method G0 B1.5 F40, the second lowest for Method G180 B1.5 F40, and the maximum for Method GOB0 F40. The ranges of the differences were-0.28 to 0.02%,-2.17 to-1.80%, and-4.92 to-4.55%, and 0.31 to-0.12%,-3.42 to-2.86%, and-7.31 to-6.92%,respectively, for 96-well and 6-well plates. The cell culture experiments proved that Method G0 B1.5 F40 was an accurate, effective, simple, and practical irradiation method. The most accurate and effective cell irradiation method should always be used, as it will reduce dose differences and instability factors and provide improved accuracy and comparability for laboratories researching cellular radiosensitivity.
基金Grant-in-Aid for Encouragement of Young Scientists from Japan Society for the Promotion of Science(to A.M.)Pancreas Research Foundation of Japan(to A.M.)
文摘AIM: Activated pancreatic stellate cells (PSCs) have been implicated in the pathogenesis of pancreatic fibrosis and inflammation. Primary PSCs can be subcultured only several times because of their limited growth potential. A continuous cell line may therefore be valuable in studying molecular mechanisms of these pancreatic disorders. The aim of this study was to establish a cell line of rat PSCs by spontaneous immortalization.METHODS: PSCs were isolated from the pancreas of male Wistar rats, and conventional subcultivation was performed repeatedly. Telomerase activity was measured using the telomere repeat amplification protocol. Activation of transcription factors was assessed by electrophoretic mobility shift assay.Activation of mitogen-activated protein (MAP) kinases was examined by Western blotting using anti-phosphospecific antibodies. Expression of cytokine-induced neutrophil chemoattractant-1 was determined by enzyme immunoassay.RESULTS: Conventional subcultivation yielded actively growing cells. One clone was obtained after limiting dilution,and designated as SIPS. This cell line has been passaged repeatedly more than 2 years, and is thus likely immortalized.SIPS cells retained morphological characteristics of primary,culture-activated PSCs. SIPS expressed α-smooth muscle actin, glial acidic fibrillary protein, vimentin, desmin, type Ⅰ collagen, fibronectin, and prolyl hydroxylases. Telomerase activity and p53 expression were negative. Proliferation of SIPS cells was serum-dependent, and stimulated with platelet-derived growth factor-BB through the activation of extracellular signal-regulated kinase. Interleukin-1β activated nuclear factor-κB, activator protein-1, and MAP kinases.Interleukin-1β induced cytokine-induced neutrophil chemoattractant-1 expression through the activation of nuclear factor-κB and MAP kinases.CONCLUSION: SIPS cells can be useful for in vitro studies of cell biology and signal transduction of PSCs.
基金Supported by the National Natural Science Foundation of China, No.39100119
文摘INTRODUCTIONCrypt epithelial cells in normal small intestineproliferate at a high speed. But they are verydifficult to culture in vitro and passage stably. A lotof studies have been done[1-16]. Some domestic labsisolated and cultured crypt cells from embryonalintestines and aseptic animal intestine, but failed.We introduced normal rat epithelial cell line-IEC-6from the USA and its living condition for stablepassage was successfully established after trials. Thecell line was testified to be the small intestinalepithelial cell by electron microscopy,immunihistochemistry and enzymatic histoch-emistry. It has been applied to some relatedresearch work[17-21]. It was found that manyfactors were involved in the culture system. Ourpresent study focuses on the culture method and theinfluencing factors on IEC-6.
基金supported by grants from the National Program on Key Basic Research Project of China(973 Program),No.2014CB542200the National Natural Science Foundation of China,No.31271284,81301570+2 种基金Program for New Century Excellent Talents in University of Ministry of Education of China,No.BMU20110270the Natural Science Foundation of Shandong Province of China,No.Y2008C18Yantai Science and Technology Development Program of China,No.2011207,2011209
文摘Preliminary animal experiments have confirmed that sensory nerve fibers promote osteoblast differentiation, but motor nerve fibers have no promotion effect. Whether sensory neurons pro- mote the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells remains unclear. No results at the cellular level have been reported. In this study, dorsal root ganglion neurons （sensory neurons） from Sprague-Dawley fetal rats were co-cultured with bone marrow mesenchymal stem cells transfected with green fluorescent protein 3 weeks after osteo- genic differentiation in vitro, while osteoblasts derived from bone marrow mesenchymal stem cells served as the control group. The rat dorsal root ganglion neurons promoted the prolifera- tion of bone marrow mesenchymal stem cell-derived osteoblasts at B and 5 days of co-culture, as observed by fluorescence microscopy. The levels of mRNAs for osteogenic differentiation-re- lated factors （including alkaline phosphatase, osteocalcin, osteopontin and bone morphogenetic protein 2） in the co-culture group were higher than those in the control group, as detected by real-time quantitative PCR. Our findings indicate that dorsal root ganglion neurons promote the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells, which pro- vides a theoretical basis for in vitro experiments aimed at constructing tissue-engineered bone.
文摘Objective\ To test the effect of hypothermia on Na+/H+ exchange, activated by shrinkage and cytoplasmic acidosis. Method\ Amiloride-sensitive Na+ influx in guinea pig red cells was traced with isotope 22Na and intracellular Na+ concentration was measured by emission flame photometry. Result\ Amiloride-sensitive Na+ influx decreased linearly as a function of temperatures (about 37℃) in shrunken cells, but increased in acidified cells. The up-regulation of acid-induced Na+/H+ exchange by elevated temperature was enhanced by hypo-osmolarity. Less sensitivity of intracellular H+ site at 41℃ may be the mechanism for the inhibition of shrinkage-induced Na+/H+ exchange by elevated temperature. Heating-mediated explosive increase in the activity of acid-induced Na+/H+ exchange may be due to enhanced extracellular Na+ sensitivity and lower intracellular pH caused by acidic metabolites. Acid-induced Na+/H+ ewxchange contributes to cytoplasmic Na+ accumulation. Conclusion\ These two modes of Na+/H+ exchange with different response to elevated temperature may play different roles in the cellular pathogenesis of heatstroke.
文摘To assess, in vitro, the cellular viability in a murine macrophage cell line J774 with 9 different orthodontic wires and to evaluate the effects of its NO production. To assess cellular viability by MTT: 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl tetrazolium bromide assay in the cell line J774 with 9 different orthodontic wires and quantify NO production by these macrophages. Cell cultures were evaluated at 24, 48 and 72 hours. There was no significant difference of the means of cellular viability between the control and the group of wires in the respective time intervals. In the comparison with the control group, there was significant difference in the NO production in groups 1, 6, and 9 at 24 hours interval. Group 8 showed significant difference in relation to the control group at final time interval. Cellular viability in all groups was higher at the final time interval than at the initial time interval. This increase was significant in the control group. In the material groups, the final mean of cellular viability at 72 hours showed no significant difference when compared with the control group. NO production in all groups was higher at the final time interval than at the initial time interval. This increase was significant in the control group. In the material groups, the final mean of NO production at 72 hours was only significant in group 8 (betatitanium) when compared with the control group.
文摘The human dermis presents an ongoing problem for regenerative medicine. Current medical management uses various acellular dermal matrices on wound sites. The challenge for scientists is to examine, then to question accepted conventional wisdom and to present new concepts. In this paper, Autologous Cell Therapy will be described by using cell culture of autologous dermal fibroblasts and their extracellular matrix as a foundation for rebuilding the dermis in conditioned wound beds. This proposal seems to create a conflict with the medical approach to keeping a wound bed “moist”.
文摘OBJECTIVE: To investigate the proliferation and differentiation of cultured corneal stem cells and determine the effect of corneal stem cells cultured on amniotic membranes on the limbal area for treating corneal burns. METHODS: The proliferation and differentiation of corneal stem cells in vitro had been examined using colony-forming efficiency and immunohistochemistry. The stem cells had been cultured on amniotic membranes and transplanted to the limbal area for treating corneal burns. RESULTS: Corneal stem cells had a high proliferation capacity in primary and first passage, cytokeratin 3 was not expressed in primary culture but partly in first passage. The stem cells could proliferate to form cell layer on an amniotic membrane. When transplanted, stem cells could survive on limbus. After transplantation, ocular inflammation resolved, the cornea re-epithelialized, the stromal opacity reduced, the superficial neovascularity was lessened and the conjunctival fornix re-established. CONCLUSIONS: Ocular surface conditions could be improved by allograft of corneal stem cells cultured on amniotic membranes.
