Hek293 cells adherent or suspension. in lysis buffer at 4 °C for cell disruption.
Hek293 cells adherent or suspension. Avoid touching the cells when replacing media.
Hek293 cells adherent or suspension. Box 4, long-term storage of cell lines in liquid nitrogen: ~1 h? TROUBLESHOOTING. The modal chromosome number was 64, occurring in 30% of cells. In addition to its ease of growth in serum-free suspension culture and its amenability to transfection, this cell line’s most important attribute is its human origin, which makes it suitable to produce biologics intended for human use. We unexpectedly found that HEK293 cells belonged to a third group Nov 14, 2023 · The SARS-CoV-2 N-expressing HEK-293 cell pools obtained (HEK-293-N-100 cells and HEK-293-N-200 cells) were used to study the degradation kinetics of the SARS-CoV-2 N protein in cell culture supernatant of adherent HEK-293 cells. 31 and 293T) or suspension (293-H, 293-F and Freestyle 293-F) cells. High Transfection Efficiency: Both cell lines exhibit high transfection efficiency, allowing for efficient introduction and expression of antibody genes. 001) higher in E1A expression levels in HEK293, 293E and 293-H cell lines compared to others, whereas E1B was Mar 24, 2015 · In this study, we examined the surface stiffness of embryonic kidney HEK293 cells in adherent and suspended states using the AFM indentation method. The genomes and the transcriptomes of these six cell lines The two predominant methods include transient transfection of suspension-adapted mammalian HEK293 cells 6–8 and the infection of Spodoptera frugiperda (Sf9) insect cells using the baculovirus expression vector system (BEVS). Transfer a small aliquot of the cell suspension to a microcentrifuge tube and dilute the cells such that the total number of cells counted will not be less than 100 or more than 1,000. 3 × 10 6 cells per well in 1 mL of DMEM/F12-FBS in triplicates in 24-well plates. The growth achieved at N stage exceeded the target viable cell density (VCD) of 2x10 6 cells/mL for transfection. host cells, and typically HEK293 cells are the cell line of choice. Direct adaptation process of HEK293 cells. 1. The University of Sheffield. Nov 10, 2019 · HEK293 cell line, derived from human embryonic kidney cells, is a well-established system for the production of LVs since they are well adapted to suspension cultures and easy to transfect . For suspension cells only, “spinoculate” by wrapping the perimeter with parafilm, placing the plate into the centrifuge with an appropriate balance, and spin the cultures at 1,200 × g at +25°C for 2 hours. Gently aspirate to mix. Oct 25, 2022 · However, their large-scale productions with Good Manufacturing Practices yield low efficiency when adherent and serum dependent HEK293 (Human Embryonic Kidney) cells are used as the host. Box 1, growth and maintenance of adherent and suspension HEK 293 cells: ~30 min. Six Feb 20, 2023 · To further look into this we here present data from 141 individual cells from a CHO-K1 cell line suspension culture and 61 single-cells from HEK293FT cells grown in adherent culture, using single-cell RNA sequencing (scRNASeq) with the Smart-seq2 (SS2) v2 protocol (Picelli et al. Serum-Free HEK Cell Culture Media has been optimized for the growth and expansion of HEK293 cells under serum-free, low protein (<75 μg/mL) conditions in suspension culture. (B) Cell volumes were measured on the second day in 5 HEK293T adherent cells were adapted to suspension cells in a serum-free medium to establish large-scale processes for functional LV production in a stirred bioreactor without micro-carriers. , HEK293-T) or in suspension (e. Table S1 provides full results of transcript levels (TPM) for all cell lines. Jang Ellen Sofie Pete P. We unexpectedly found that HEK293 cells belonged to a third group Jul 2, 2021 · The HEK293 cell line has earned its place as a producer of biotherapeutics. Box 3, FACS to enrich subpopulations of transduced cells: ~1 d. Frozen cells are supplied in and may be thawed directly into FreeStyle™ 293 Expression Medium. To transition to serum-free suspension culture, cells were detached and centrifuged at 200 × g for 5 minutes and resuspended in new, serum-free medium at a concentration of 4 – 6 × 105 cells/mL. FreeStyle 293 cells) in suspension. 05, **p < 0. Sep 6, 2022 · A) Bright-field images of adherent and suspension HEK293 cells after completing serum-free medium cultures. This is a complete media that will support growth of a wide variety of HEK293 cells without further supplementation. We further investigated the impact of adaptation procedures, serum-free medium, and culture modes on cell growth and intracellular metabolite Dec 13, 2021 · Early viral vector production was challenging as production was performed by the transient transfection of adherent HEK293 cells in serum-containing media. Mar 21, 2016 · Growth and maintenance of adherent and suspension HEK293 cells. fection of adherent HEK293 cells has been the predominant platform [14,15]. Large-scale production of current Good Manufacturing Practices-grade lentiviral vectors is limited because of the adherent, serum-dependent nature of HEK293T cells used in the manufacturing process. The cell lines can be divided into either adherent (HEK293, 293E and 293T) or suspension (293-H, 293-F and Freestyle 293-F) cells. Dec 14, 2015 · Strategy 2. While, for example, HEK293 cells have been adapted to grow in suspension Malm M, Saghaleyni R, Lundqvist M et al. This transition to suspension culture simplifies subcultivation, enhances scalability, and reduces production costs, making it an advantageous approach The impact of serum-free culture on HEK293 cells: From the establishment of suspension and adherent serum-free adaptation cultures to the investigation of growth and metabolic profiles. It was developed from the HEK-293 cell line with the expression of a mutant SV40 large T antigen [5]. The adaptation of the HEK293 cell line to suspension culture resulted in robust growth and achieved similar or improved maximum cell density compared to adherent cultures. 01 and ***p < 0. M. Mar 2, 2021 · A Welch two sample T-test showed significantly (*p < 0. Equipment setup Shimadzu Prominence high-performance liquid chromatography (HPLC) system. in lysis buffer at 4 °C for cell disruption. Methods. 14-17 h for CHO), and product yield in stable What is the HEK293 Cell Size? HEK293 cell diameter is typically between 11 and 15µm, however, the diameter of cells is dependent upon their culturing conditions. Graham's original HEK293 cells grew in adherent culture, which means they stick to the bottom of a cell culture dish. To optimize large Aug 22, 2018 · It was established that cell deformability undergoes significant alteration within the first hours of exposure to Pluronic F-68, even at concentrations as low as 0. Figure 2. 5L to 3. Dec 18, 2023 · Cell suspensions are used in various applications in biological research, biotechnology, and medicine, including cell counting, cell viability assays, flow cytometry, cell sorting, and the production of recombinant proteins or viral vectors. The cells were obtained from a single, aborted or miscarried fetus, the precise origin of which is unclear. From the lysate, 20 µL was transferred to a black frame white well 96-well plate for luminescence measurement after the addition of substrate in buffer ( Appendix A. , HEK293-F). In this protocol, we developed a simple scalable AAV production protocol using serum-free-media-adapted HEK293T suspension cells and VirusGEN transfection reagent. One commonly used AAV production platform for pre-clin-ical research is based on the plasmid-based transfection of adherent HEK293 cells grown in serum-containing media. Sartorius offers four high-performing suspension HEK media that are designed for use with the high variability of HEK293 cell lines, the versatility of adeno-associated virus and the complexity of viral vector processes. Prepared ready-to-use, with no supplementation required. Other cytokines and growth factors may be supplemented Volume 110, 15 June 2016, Pages 17-26. e. HEK 293 cells were generated in 1973 by transfection of cultures of normal human embryonic kidney cells with sheared adenovirus 5 DNA in Alex van der Eb 's laboratory in Leiden, the Netherlands. Overall, material cost are about 40% lower and producing AAV in suspension-adapted HEK 293 cells takes only one fifth of the time compared to adherent HEK 293 culture. Jan 29, 2020 · Request PDF | Evolution from adherent to suspension – systems biology of HEK293 cell line development | The need for new safe and efficacious therapies has led to an increased focus on biologics Cell culture of HEK293T suspension cells. To increase production efficiency, HEK293 cells are adapted to grow in suspension using commercially available and chemically defined serum-free mediums. Dispensing of this step greatly simplifies the process. For all experiments, the cells were cultured in Irvine BalanCD HEK293 medium supplemented with L-glutamine, Pluronic F-68, and glucose General information. Make adherent cells come off by pipetting media up and down on them directly until all are off or use a cell scraper. Viable cell density and viability measurements during the direct adaptation process. These transfection conditions increased cell survival, allowing the selection of stable cell pools, which was otherwise not possible with standard procedures in suspension. For adherent cells, place the plate into the CO 2 incubator and grow cells under standard conditions for 16-24 hours. 5% w/v for cells in the suspension without serum. They are a hardy, semi-adherent, low-maintenance cell line and divide rapidly, doubling about every 36 hours. The transfection efficiency upon polyethyleneimine (PEI)-mediated transient gene delivery into HEK-293 cells cultured in suspension was investigated to understand the effect of different culture and transfection conditions as well as the significance of the culture age and the quality of the cell line used. Adapting cells so that they are suitable for growth in suspension Centrifuge at 300 x g for 5 minutes and plate 1 x 10 6 cells in a 60 mm dish (or 2 x 10 6 in a 100 mm dish). There are two basic systems for growing cells in culture, as monolayers on an artificial substrate (i. The FreeStyle™ 293-F cell line is supplied in a vial containing 1 mL of cells at 1 x 10 7 viable cells/mL in 90% FreeStyle™ 293 Expression Medium Feb 2, 2024 · Most laboratories commonly use adherent HEK293 or HEK293T cells as producer cells because transfection of adherent cells is more efficient than that of suspension cells. 4 rAAV vectors produced in both systems have been shown to have comparable biological activity. Suspension culture in serum-free medium offers advantages in large scale manufacturing. HEK293 cells are traditionally grown in adherent, monolayer cultures. 32 and the transcriptomes of these six cell lines were sequenced using Illumina HiS-. Contains no human or animal-origin products. Bruheim Jan 13, 2022 · HEK293 cells, also known as HEK 293 cells, HEK-293 cells or less precisely as HEK cells, are human embryonic kidney cells which grow in tissue culture and represent a distinct immortalised cell line. Changes in gene expression between adherent and suspension derivatives highlighted switching in cholesterol biosynthesis and expression of five key genes (RARG FreeStyle™ 293-F cells are adapted to suspension culture in FreeStyle™ 293 Expression Medium. Avoid touching the cells when replacing media. Whereas the yield obtained with adherent cells is generally limited by the cell culture surface area, suspension-grown cell lines show better and more straightforward scalability. Troubleshooting guidance can be found in Nov 9, 2021 · On the other hand, switching to serum free, suspension platform is not always a viable solution and is far from being a failure free endeavor. HEK293 cells are typically maintained in adherent cultures, but far greater culture volumes are possible with cells floating free in suspension. Direct: add cells suspension Sep 27, 2021 · There are many advantages of using HEK293 cells. Recent advancements in transient transfection technologies, cell line and media development have demonstrated that viral vector production can be performed using serum-free suspension-based The suspension HEK293 were grown in adherent conditions, with 20% FBS supplementation. The Human Embryonic Kidney 293T (HEK-293T) cell line is widely used for the production of recombinant proteins and lentiviral vectors, since it is easy to transfect and can be adapted to serum-free suspension culture [3,4]. We believe that the Expi293F cell line is a significant improvement over traditional HEK293 cell lines due to its robust growth under suspension culture conditions as well as its property of stable expression in addition to transient expression. HEK293 stable cell line by random integration. Both protocols are Jan 30, 2020 · Changes in gene copy and expression between industrial progeny cell lines and the original HEK293 were associated with cellular component organization, cell motility and cell adhesion. 2 ). ATCC CRL-1573 (HEK293) cells were progressively adapted from an anchorage-dependent growth mode to a suspension mode. parental HEK293 master cell bank (MCB) was performed Characteristics. Apr 16, 2023 · The data indicate that HEK293 cells are primarily used for the commercial production of gene and chimeric antigen receptor (CAR) T cell therapeutics. g. and Tan et al. The presence of sodium heparin in the medium formulation reduced cell clumping dramatically in suspension cul … Feb 5, 2024 · Although suspension cells are easier to handle and scale up compared to adherent cells, their use in virus production is hampered by poor transfection efficiency. Ensure that cell Oct 1, 2021 · HEK293 cell lines are used for the production of recombinant proteins, virus-like particles and viral vectors. Jun 18, 2013 · The transfection efficiency upon polyethyleneimine (PEI)-mediated transient gene delivery into HEK-293 cells cultured in suspension was investigated to understand the effect of different culture and transfection conditions as well as the significance of the culture age and the quality of the cell line used. The cells were seeded at 0. To 1 ml of the diluted cell suspension, add 100 μl Trypan Blue Stain (0. HEK293-F6 suspension cells were transfected (PEI-DNA method) and positive cell pool was selected by puromycin addition to the media. Transitioning cells to suspension culture is the first step in the journey towards a scalable AAV production process. 1-2 × 10 7 for CHO) , cultivation time (HEK293 doubling time 33 h vs. Nov 3, 2015 · AAV was found to be secreted by an unknown mechanism into cell culture media using adherent HEK293 cells and harvesting rAAV from the media at a late time-point post-transfection. T-150 flask (Corning, 150 cm 2) Five chamber cell stack (CF5, Corning, 3180 cm 2) Heat-inactivated FBS (HI-FBS) Lentiviral vectors are increasingly utilized in cell and gene therapy applications because they efficiently transduce target cells such as hematopoietic stem cells and T cells. They can be utilized for both transient and stable expression, can be cultured in suspension or as a monolayer, are easy to transfect (and can be transfected via a variety of methods Dec 13, 2021 · Early viral vector production was challenging as production was performed by the transient transfection of adherent HEK293 cells in serum-containing media. Adeno-associated virus (AAV) has shown great promise as a gene therapy vector in multiple aspects of preclinical and clinical applications. 5L, more cells can be achieved in Corning 5L Erlenmeyer flasks with-out decreasing the viability. Nov 24, 2011 · In contrast, suspension-grown HEK293 cells in combination with PEI-mediated transfection were used in this work. . [3] [2] The cells were cultured by van der fection of adherent HEK293 cells has been the predominant platform [14,15]. The following is based on the parent cell line ( ATCC CRL-1573 ): This is a hypotriploid human cell line. Best of luck! Ceren Pekşen I Jul 11, 2023 · Adherent and Suspension Growth: Both HEK and CHO cells can grow as adherent cells in monolayer cultures, or within suspension culture for improved scalability. Apr 16, 2023 · HEK293 is a widely used cell line in the fields of research and industry. Dec 13, 2021 · Early viral vector production was challenging as production was performed by the transient transfection of adherent HEK293 cells in serum-containing media. Sci. , 2012). Jul 17, 2007 · Moreover, we have grown the HEK 293 suspension cells in reusable glass bottles, whereas adherent cells require specially treated and expensive plastic surfaces to grow. The embryonic human kidney (HEK) 293AAV cell line (Cell Biolabs, #AAV-100), which was cloned and selected from parental 293 cell line (transformed with human adenovirus type 5 DNA), was used to produce AAV in adherent culture. Figure 3. 293 cells may be subcultured either in suspension culture or as adherent monolayers depending upon the application. HEK293 in vaccine production Nov 4, 2020 · Changes in gene copy and expression between industrial progeny cell lines and the original HEK293 were associated with cellular component organization, cell motility and cell adhesion. In this application note, we demonstrate the performance of PEI is just as good as Lipofectamine 2000 for HEK293 cells, and yes, you should linearize your plasmid first if genomic integration is the goal. Once your cells are no longer dependent on a surface for growth, scale-up Figure 4 shows the total cell yield of HEK-293F at day 8 with three different fill volumes. The portfolio also Oct 1, 2023 · To generate stable suspension cells expressing the RBD-SrtA, a single two plasmids transfection was performed, with hygromycin selection. We adapted adherent HEK293T cells to suspension culture in our HyClone™ transfection medium, HyCell™ Transfx-H. This production method is dif ficult to scale up, labor intensive, and incompatible with current GMP-compliant manufacturing practices. Nov 14, 2023 · Watch the video or view the poster to:Learn how to cultivate HEK293 in suspension in stirred-tank bioreactorsGet hands-on advice to improve a suspension cell cultureUnderstand MSC-derived exosome production, isolation, and quantification Jun 1, 2023 · Generally, adaptation of adherent HEK293 cells into chemically defined suspension media can occur quite readily, notes Zmuda—as long as care is taken to perform the adaptation at relatively high cell densities to not stress the cells during the process. Scale bars indicate 100um. Adherent processes require cells to be disassociated from their growth surface for routine maintenance, scale-up, and counting. HEK293 in vaccine production This includes cell dissociation, counting cells, determining optimal seeding density and preparing new culture vessels for passaged cells. HEK293 cells were cultured in X medium with X% FBS. HEK293 cells are transformed cell line with adenovirus ( Graham et al. , adherent culture) or free-floating in the culture medium (suspension culture). transient transfection technology and mammalian HEK293 cells. Cell growth of VirusExpress ® 293 AAV cells in a Mobius ® 50 L Bioreactor at 10 L and 40 L working volume. However, suspension cells are easier to handle and scale up, which is of particular interest to Good Manufacturing Practice (GMP) facilities. A detailed analysis of different HEK293 cell lines can be found in the studies by Malm et al. However, their large-scale productions with Good Manufacturing Practices yield low efficiency when adherent and serum dependent HEK293 (Human Embryonic Kidney) cells are used as the host. At the present time, the growth and production properties of the HEK293 cell line Sep 6, 2022 · Adherent HEK293 cells were adapted to grow in a serum-free medium (Freestyle 293 expansion medium) following two different procedures and established serum-free suspension cultures from these cells. To begin, an adherent HEK293 cell line from our GMP facil-ity’s qualified master cell bank (derived from a clone previously selected for high transfection efficiency and rAAV production) was adapted to grow in serum-free suspension conditions as described in the methods section. The aim of this research was to use particle image velocimetry validated computational fluid dynamics (CFD) to determine the hydrodynamic stress in both shake flasks, with and without baffles, and in stirred Minifors 2 bioreactors to evaluate its effect Oct 6, 2015 · Derivation of suspension HEK293 cells from an adherent HEK293 quali-fied master cell bank. 1 to 3: (1) HEK 293 cell line A with Xell medium 1 (w/o FCS), (2) HEK 293 cell line B with Xell medium 2 (w/o FCS), (3) HEK 293 cell line B with Xell medium 3 (w/o FCS). 2 to 0. 293T cells, but not the parental HEK293 line, are Adherent HEK293T cells (ideally AAV-293T clones) *Pro-Tip* While adherent, these cells are very loosely attached to the dish surface and should be handled carefully. The bioreactor is utilized for cell expansion (N-1 stage) prior to the virus production stage (N stage). CapPCV2 gene sequence was codon optimized for mammalian cell expression. Jul 8, 2021 · A suspension process has the advantage of being easier to operate than an adherent one. e derivation of the suspension cell line from the . Record the dilution factor. This objective was achieved with three recombinant HEK-293 cell lines expressing 5HT receptors, which were successfully adapted for robust growth in suspension culture using a new serum-free medium, Ex-Cell 520 (JRH Biosciences). Here we describe the optimized transfection of HEK 293T cells in both culture formats. The genomes. HEK293 Genome and Chromosome Number Ian R Wilkinson. Growth and maintenance of Ric15(Gntl(-)) suspension cells Nov 14, 2019 · We sequenced poly(A)-selected RNAs isolated from all four GLUL −/− cell lines and the wildtype HEK293 cells from adherent and suspension cultures during exponential phase, and obtained between 19 to 147 million paired-end reads for each sample. 4 Production platforms The biggest limitation to this approach is scalability. Feb 1, 2016 · This work has successfully adapted an adherent HEK293 cell line from a qualified clinical master cell bank to grow in animal component-free suspension conditions in shaker flasks and WAVE bioreactors that allows for rapid and scalable rAAV production. 8°C-- inoculation density: 3x105 cells/mL - fill volume: 15 mL, inoculum volume: 2 mL 5) Use 10mL pipette to suspend cells. The HEK293T suspension cells were routinely passaged/subcultured three times per week at an inoculum VCD of 0. 6) Put all of the media from all 5 flasks into a baffled flask 7) Put the 100mL of cells in shaker for 3 or 4 days. The HEK 293F cell line (Gibco, Buffalo, NY) that was adapted to serum-free medium was used to produce AAV in suspensive An optimized, serum-free, protein-free formulation designed to support the high-density culture and transfection of 293 cells (e. May 20, 2020 · Graham's initial HEK293 cells were grown in adherent culture, meaning they were attached to the bottom of a cell culture dish. Human Embryonic Kidney 293 (HEK293) cells were adapted into a serum-free suspension medium through steps of gradual serum weaning for the production of adenoviral (AdV) gene therapy vectors. Recent work has generated molecular (systems level) characterisation of HEK293 variants that has enabled re-engineering of the cells towards enhanced use for manufacture-scale production of recombinant biopharmaceuticals (assessment of ‘safe harbours’ for gene insertion The HEK293 cell line has earned its place as a producer of biotherapeutics. The obtained sequence was cloned into pIRESpuro3 bicistronicvector . 32,33,34 This concept was explored using the suspension HEK293 production system described in this report with the exception that media was harvested and replaced at Jan 30, 2020 · The cell lines can be divided into either adherent (HEK293, 293E. 33 eq. Viable cell density and viability of HEK-293F cells cultured in Jun 30, 2021 · Human Embryonic Kidney 293 cells can be both grown as adherent or suspension but usually, these cells are cultured as an adherent monolayer. Processes no. To keep HEK 293 cells alive, a high-glucose cell culture media is used. Evolution from adherent to suspension: systems biology of HEK293 cell line development. The medium is not recommended for adherent 293 cell culture. Cells were maintained at 37 °C and 5% CO2 at a shaking frequency of 125 – 185 rpm Jul 2, 2021 · Yet, despite these significant improvements in recombinant protein and virus production, this cell line continues to lag behind the CHO cell line in growth capacity (Maximal cell density of 3-5 × 10 6 /mL for HEK293 vs. To create a cell suspension of adherent cells, such as HEK293 cells, the following steps can be performed: Growth in gene therapies and viral vector vaccines has increased the demand for robust HEK293 cell lines. The adaptation process was cell line dependent and the time scale of this process varied between 4 and 8 weeks. HEK293 cells can grow adherent (e. Recent advancements in transient transfection technologies, cell line and media development have demonstrated that viral vector production can be performed using serum-free suspension-based Jun 6, 2019 · HEK293 cells were transfected using calcium phosphate transfection for adherent AAV293 cells grown in cell factories and PEI for HEKExpress suspension cells grown in Tubespin 600 bioreactors. The results showed that 293 T suspension was successfully cultivated in F media (293 CD05 medium and SMM293-TII with 1:1 volume ratio), and the cells This set of cell lines includes the parental HEK293 as well as five additional cell lines that have all been clonally derived from parental HEK293 cells. 4%) solution. It is assumed that these cells are sensitive to hydrodynamic stress. Subculture cells 3–5 days post thaw or when cultures reach 60– 80% confluence (adherent monolayer), or a viable cell density of 2 × 106–3× 106 viable cells/mL (suspension). Recent advancements in transient transfection technologies, cell line and media development have demonstrated that viral vector production can be performed using serum-free suspension-based Apr 16, 2020 · In Oxford, scientists at the University's Clinical Biomanufacturing Facility adapted suspension HEK293 cells to serum free suspension culture. Box 2, endpoint dilution assay to determine functional lentiviral titer: ~4 d. If this does not work, try adding DNase to your media. Although HEK cells and Chinese hamster ovary cells (CHO cells) are both used to create antibodies and for therapeutic protein production, they Sep 14, 2022 · For both adherent cells and suspension cells, the cells were incubated for 20 min. Adherent HEK cells do not show the stiffening trend as observed for Jurkat cells and RBCs. Culture cells as described in Box 1. 3 × 10 6 cells/mL. A total of 5 × 10 8 cells (500 mL) per batch were transfected and harvested 72 hr post-transfection. , 1977) and they show cancer-like behavior in tissue culture. Changes in gene expression between adherent and suspension derivatives highlighted switching in cholesterol biosynthesis and expression of five key genes (RARG Feb 20, 2022 · The transient transfection of mammalian cells is a rapid and versatile platform for the manufacture of recombinant proteins, but industrial processes depend on reliable scalability and efficient conversion from adherent to suspension cell cultures. Hek cells can be adapted to suspension culture using a number of techniques: The most common ones we have used are: 1. To develop a perfusion platform, TFF and ATF bioreactor experiments were designed to optimize perfusion method with serum-free and suspension adapted clonal HEK293 cells. The suspension HEK293 were grown in adherent conditions, with 20% FBS supplementation. , 2014) and SMART-Seq (Ramsköld et al. If the volume is increased from 2. Many On the other hand, switching to serum free, suspension platform is not always a viable solution and is far from being a failure free endeavor. Oct 25, 2021 · (A) Cell growth kinetic and (B) Glucose and lactate profile of adherent serum-supplemented HEK-293T cells cultured in a 24-well plate with an initial cell density of 7x10 3 cells/mL for 8 days (n Jul 30, 2015 · In this study, we examined the surface stiffness of embryonic kidney HEK293 cells in adherent and suspended states using the AFM indentation method. Cells grown in an adherent monolayer culture will appear flatter and have a larger diameter than cells in suspension culture, which will be spheroid. Adherent HEK293T cells have been adapted to suspension culture in serum-free - conditions optimal culture medium was CD293 + 4mM Glutamax (Gibco) as determined - in a previous study Setpoints Ambr ® 15 culture using sparged vessels: - bioreactor temperature: 36. hfrbrtpfiwxxfgcmldry