Misunderstandings often encountered in primary cell culture
A primary culture cell refers to a cell cultured immediately after being taken out of the body. The first generation cells cultured and the cells within 10 generations are collectively referred to as primary cell culture. Primary cells are not only widely used in molecular, cell biology and biomedical basic research, such as proteomics, genomics, cell line (system) research, DNA, RNA and genetic research, but also for today's popular organisms. Pharmaceutical industry such as drug screening, drug metabolism and toxicology research, cancer drug research, etc. Primary cells have an irreplaceable role in the field of biomedicine and have broad market prospects.
The culture of primary cells refers to culture immediately after removing cells, tissues and organs directly from the body. Therefore, strictly speaking, it refers to the culture before successful passage. At this time, the cells maintain the basic properties of the original cells, and if they are normal cells, the diploid number is retained. However, in practice, cultured cells from the first to the tenth generation are generally referred to as primary cell culture.
The following is a small series to help you correct the problems that arise in primary cell culture.
Error 1: A small tube of primary cells is thawed in a water bath for a while
Correction 1: Primary cells are very sensitive to the thawing process, so it is important to place the vial in a 37 ° C water bath and keep it and gently rotate until the contents have just thawed. The vial should then be removed from the water bath immediately and transferred to a sterile workstation. Make sure your flask is ready before thawing so that it can be used immediately to inoculate cells and placed in an incubator.
Error 2: Centrifuge the primary cells directly after thawing the match vial
Correction 2: We do not recommend centrifuging the cells after thawing because the centrifugation procedure is more harmful than a small amount of DMSO residue. Remember to change the medium the next day after restoring the primary cells to remove any residual DMSO.
Mistake 3: Allowing primary cells to become too fused
Correction 3: Primary cells can become senescent when grown to 100% confluence. Remember that primary cells are not 100% pure, so it is important to minimize the growth of contaminating cells. We recommend subculture of primary cells when the cells reach 90-95% confluency.
Mistake 4: Excessive trypsin digestion when passaged primary cells
Correction 4: When cells were passaged, low concentrations of trypsin were used and the cells were closely monitored under a microscope. In addition, remember to completely neutralize trypsin in the cells after trypsinization, as any active trypsin will damage the cells.
Mistake 5: Primary cells can be easily re-frozen
Correction 5: Usually we do not recommend re-frozen primary cells as this can promote cellular senescence and/or cause functional changes. Primary cells are very sensitive and re-frozen can cause cell death or damage.
Mistake 6: Primary cells can proliferate indefinitely
Correction 6: Unlike cell lines, primary cells have limited amplification capabilities. We recommend using primary cells as early as possible to experiment to prevent genetic drift. In addition, if you are using a cell type that is difficult to add value, you should closely monitor the cell morphology, as a small number of mixed cells (such as fibroblasts) may grow in large quantities over time to become the main cell.
Beijing Baiou Bowei Biotechnology Co., Ltd.'s China Microbial Species Enquiry Network provides services such as microbial strain preservation, sequencing, and purchase. It is a service platform for the China Microbial Culture Collection, and is a collection of microbial strains, strains, and ATCC strains. A large-scale microbiological inquiry website that integrates cells and culture media, and a microbial culture collection center with equipment and technology! Welcome customers to inquire!
A primary culture cell refers to a cell cultured immediately after being taken out of the body. The first generation cells cultured and the cells within 10 generations are collectively referred to as primary cell culture. Primary cells are not only widely used in molecular, cell biology and biomedical basic research, such as proteomics, genomics, cell line (system) research, DNA, RNA and genetic research, but also for today's popular organisms. Pharmaceutical industry such as drug screening, drug metabolism and toxicology research, cancer drug research, etc. Primary cells have an irreplaceable role in the field of biomedicine and have broad market prospects.
The culture of primary cells refers to culture immediately after removing cells, tissues and organs directly from the body. Therefore, strictly speaking, it refers to the culture before successful passage. At this time, the cells maintain the basic properties of the original cells, and if they are normal cells, the diploid number is retained. However, in practice, cultured cells from the first to the tenth generation are generally referred to as primary cell culture.
The following is a small series to help you correct the problems that arise in primary cell culture.
Error 1: A small tube of primary cells is thawed in a water bath for a while
Correction 1: Primary cells are very sensitive to the thawing process, so it is important to place the vial in a 37 ° C water bath and keep it and gently rotate until the contents have just thawed. The vial should then be removed from the water bath immediately and transferred to a sterile workstation. Make sure your flask is ready before thawing so that it can be used immediately to inoculate cells and placed in an incubator.
Error 2: Centrifuge the primary cells directly after thawing the match vial
Correction 2: We do not recommend centrifuging the cells after thawing because the centrifugation procedure is more harmful than a small amount of DMSO residue. Remember to change the medium the next day after restoring the primary cells to remove any residual DMSO.
Mistake 3: Allowing primary cells to become too fused
Correction 3: Primary cells can become senescent when grown to 100% confluence. Remember that primary cells are not 100% pure, so it is important to minimize the growth of contaminating cells. We recommend subculture of primary cells when the cells reach 90-95% confluency.
Mistake 4: Excessive trypsin digestion when passaged primary cells
Correction 4: When cells were passaged, low concentrations of trypsin were used and the cells were closely monitored under a microscope. In addition, remember to completely neutralize trypsin in the cells after trypsinization, as any active trypsin will damage the cells.
Mistake 5: Primary cells can be easily re-frozen
Correction 5: Usually we do not recommend re-frozen primary cells as this can promote cellular senescence and/or cause functional changes. Primary cells are very sensitive and re-frozen can cause cell death or damage.
Mistake 6: Primary cells can proliferate indefinitely
Correction 6: Unlike cell lines, primary cells have limited amplification capabilities. We recommend using primary cells as early as possible to experiment to prevent genetic drift. In addition, if you are using a cell type that is difficult to add value, you should closely monitor the cell morphology, as a small number of mixed cells (such as fibroblasts) may grow in large quantities over time to become the main cell.
Beijing Baiou Bowei Biotechnology Co., Ltd.'s China Microbial Species Enquiry Network provides services such as microbial strain preservation, sequencing, and purchase. It is a service platform for the China Microbial Culture Collection, and is a collection of microbial strains, strains, and ATCC strains. A large-scale microbiological inquiry website that integrates cells and culture media, and a microbial culture collection center with equipment and technology! Welcome customers to inquire!
2004-2021 New Laser Distance Sensor
Are you looking for a distance photoelectric sensor with mm high accuracy? Yes, JRT manufactured the industrial laser lidar sensor since 2004. There are different models of high accuracy sensor, high frequency distance sensors, ip54 ip65 ip67 waterproof rating etc., you can choose. The laser measuring modules, some good for outdoor using, some can be connected to your Arduino,
Also, Analog Output, RS485, USB cable is optional. If you have any other requirements of our accuracy distance sensor, please fell free to contact us
Chengdu JRT Main Ranging Modules: Industrial Laser Distance Sensors, Laser Rangefinder Sensor , Tof LiDAR Sensors.
smart devices,photoelectric sensor,cmos sensor,distance laser sensor ,laser lidar
Chengdu JRT Meter Technology Co., Ltd , http://www.rangingsensor.com