AIM:The generation and characterization of a human embryonic stem cell (hESC) line stably expressing red fluorescent mCherry protein. METHODS:Lentiviral transduction of a ubiquitously-expressed human EF-1α promoter d...AIM:The generation and characterization of a human embryonic stem cell (hESC) line stably expressing red fluorescent mCherry protein. METHODS:Lentiviral transduction of a ubiquitously-expressed human EF-1α promoter driven mCherry transgene was performed in MEL2 hESC. Red fluore-scence was assessed by immunofluorescence and flow cytometry. Pluripotency of stably transduced hESC was determined by immunofluorescent pluripotency marker expression, flow cytometry, teratoma assays andembryoid body-based differentiation followed by reverse transcriptase-polymerase chain reaction. Quantification of cell motility and survival was performed with time lapse microscopy. RESULTS:Constitutively fluorescently-labeled hESCs are useful tools for facile in vitro and in vivo tracking of survival, motility and cell spreading on various surfaces before and after differentiation. Here we describe the generation and characterization of a hESC line (MEL2) stably expressing red fluorescent protein, mCherry. This line was generated by random integration of a fluorescent protein-expressing cassette, driven by the ubiquitously-expressed human EF-1α promoter. Stably transfected MEL2-mCherry hESC were shown to express pluripo-tency markers in the nucleus (POU5F1/OCT4, NANOG and SOX2) and on the cell surface (SSEA4, TRA1-60 and TG30/CD9) and were shown to maintain a normal karyotype in long-term (for at least 35 passages) culture. MEL2-mCherry hESC further readily differentiated into representative cell types of the three germ layers in embryoid body and teratoma based assays and, importantly, maintained robust mCherry expression throughout differentiation. The cell line was next adapted to singlecell passaging, rendering it compatible with numerous bioengineering applications such as measurement of cell motility and cell spreading on various protein modified surfaces, quantification of cell attachment to nanoparticles and rapid estimation of cell survival. CONCLUSION:The MEL2-mCherry hESC line conforms to the criteria of bona fide pluripotent stem cells and maintains red fluorescence throughout differentiation, making it a useful tool for bioengineering and in vivo tracking experiments.展开更多
Zinc and its compounds, alloys and composites play an important role in the modern day world and find application in almost every aspect that can improve the quality of our lives. This ranges from supplements and phar...Zinc and its compounds, alloys and composites play an important role in the modern day world and find application in almost every aspect that can improve the quality of our lives. This ranges from supplements and pharmaceuticals that are meant to improve our health and wellbeing to additives meant to guard or reduce corrosion in metals. However, over the past several years, a new area of technology has been garnering a great deal of attention and has made use of zinc and its compounds. This is with reference to paper-based microfluidic technology that offers several advantages and that keeps expanding in the amount of applications it covers. In this paper, a review is offered for the applications that have used zinc or zinc compounds in paper-based microfluidic devices.展开更多
Chronic wounds are wounds that are difficult to heal or do not follow the normal healing process.These include pressure ulcers,diabetic ulcers,venous ulcers,and arterial incomplete ulcers.Unlike acute wounds,chronic w...Chronic wounds are wounds that are difficult to heal or do not follow the normal healing process.These include pressure ulcers,diabetic ulcers,venous ulcers,and arterial incomplete ulcers.Unlike acute wounds,chronic wounds are often difficult to heal or even do not heal.Its pathogenesis involves many factors;bacterial infection is the main cause of chronic wound.With the increase in population aging,the incidence of chronic wounds has become a critical issue in the current medical and health field.Management of chronic wounds is faced with the problems of long treatment time,difficulty,high cost,repeated attacks and high disability rate,which seriously threaten patients'ability to take care of themselves in normal life and cause a heavy burden to individuals,families and society.Over the past few years,the development of antibacterial hydrogels for the treatment of bacterial infections has received a lot of attention.Since antibacterial hydrogels not only have the mechanical properties of hydrogels,high biocompatibility and adjustable functional structure,but also have excellent antibacterial properties,they may be an ideal dressing to solve the problem of chronic wound healing.This article introduces the types of chronic wounds,their healing characteristics,and the challenges faced in treating chronic wounds.It classifies antimicrobial hydrogels based on their antimicrobial modes and further discusses the advancements in smart antimicrobial hydrogels along with the benefits and obstacles of using antimicrobial hydrogels in the treatment of chronic wounds.This article also explores the development directions of antimicrobial hydrogels for chronic wound management.展开更多
基金the Australian Stem CellCentre (ASCC), UQ AIBN Challenge Grant Scheme for financial supportthe ASCC’s StemCore pluripotent stem cell core facility for the provision of hES cell lines and logistical support
文摘AIM:The generation and characterization of a human embryonic stem cell (hESC) line stably expressing red fluorescent mCherry protein. METHODS:Lentiviral transduction of a ubiquitously-expressed human EF-1α promoter driven mCherry transgene was performed in MEL2 hESC. Red fluore-scence was assessed by immunofluorescence and flow cytometry. Pluripotency of stably transduced hESC was determined by immunofluorescent pluripotency marker expression, flow cytometry, teratoma assays andembryoid body-based differentiation followed by reverse transcriptase-polymerase chain reaction. Quantification of cell motility and survival was performed with time lapse microscopy. RESULTS:Constitutively fluorescently-labeled hESCs are useful tools for facile in vitro and in vivo tracking of survival, motility and cell spreading on various surfaces before and after differentiation. Here we describe the generation and characterization of a hESC line (MEL2) stably expressing red fluorescent protein, mCherry. This line was generated by random integration of a fluorescent protein-expressing cassette, driven by the ubiquitously-expressed human EF-1α promoter. Stably transfected MEL2-mCherry hESC were shown to express pluripo-tency markers in the nucleus (POU5F1/OCT4, NANOG and SOX2) and on the cell surface (SSEA4, TRA1-60 and TG30/CD9) and were shown to maintain a normal karyotype in long-term (for at least 35 passages) culture. MEL2-mCherry hESC further readily differentiated into representative cell types of the three germ layers in embryoid body and teratoma based assays and, importantly, maintained robust mCherry expression throughout differentiation. The cell line was next adapted to singlecell passaging, rendering it compatible with numerous bioengineering applications such as measurement of cell motility and cell spreading on various protein modified surfaces, quantification of cell attachment to nanoparticles and rapid estimation of cell survival. CONCLUSION:The MEL2-mCherry hESC line conforms to the criteria of bona fide pluripotent stem cells and maintains red fluorescence throughout differentiation, making it a useful tool for bioengineering and in vivo tracking experiments.
文摘Zinc and its compounds, alloys and composites play an important role in the modern day world and find application in almost every aspect that can improve the quality of our lives. This ranges from supplements and pharmaceuticals that are meant to improve our health and wellbeing to additives meant to guard or reduce corrosion in metals. However, over the past several years, a new area of technology has been garnering a great deal of attention and has made use of zinc and its compounds. This is with reference to paper-based microfluidic technology that offers several advantages and that keeps expanding in the amount of applications it covers. In this paper, a review is offered for the applications that have used zinc or zinc compounds in paper-based microfluidic devices.
基金supported by grants from the National Key Research and Development Program of China(No.2023YFA0915200,2023YFA0915204)the equipment research and development projects of the Chinese Academy of Sciences(PTYQ2024YZ0010,PTYQ2024BJ0007)the Science and Technology Commission of Shanghai Municipality Project(XTCX-KJ-2024-038).
文摘Chronic wounds are wounds that are difficult to heal or do not follow the normal healing process.These include pressure ulcers,diabetic ulcers,venous ulcers,and arterial incomplete ulcers.Unlike acute wounds,chronic wounds are often difficult to heal or even do not heal.Its pathogenesis involves many factors;bacterial infection is the main cause of chronic wound.With the increase in population aging,the incidence of chronic wounds has become a critical issue in the current medical and health field.Management of chronic wounds is faced with the problems of long treatment time,difficulty,high cost,repeated attacks and high disability rate,which seriously threaten patients'ability to take care of themselves in normal life and cause a heavy burden to individuals,families and society.Over the past few years,the development of antibacterial hydrogels for the treatment of bacterial infections has received a lot of attention.Since antibacterial hydrogels not only have the mechanical properties of hydrogels,high biocompatibility and adjustable functional structure,but also have excellent antibacterial properties,they may be an ideal dressing to solve the problem of chronic wound healing.This article introduces the types of chronic wounds,their healing characteristics,and the challenges faced in treating chronic wounds.It classifies antimicrobial hydrogels based on their antimicrobial modes and further discusses the advancements in smart antimicrobial hydrogels along with the benefits and obstacles of using antimicrobial hydrogels in the treatment of chronic wounds.This article also explores the development directions of antimicrobial hydrogels for chronic wound management.
