High yield tissue dispersing method for cell culture

1980 ◽  
Vol 6 (3-4) ◽  
pp. 97-100 ◽  
Author(s):  
Héctor Montes Oca
Keyword(s):  
Cell Culture ◽  
High Yield

scholarly journals Defined MSC exosome with high yield and purity to improve regenerative activity

2021 ◽  
Vol 12 ◽  
pp. 204173142110086
Author(s):  
Jun Yong Kim ◽  
Won-Kyu Rhim ◽  
Yong-In Yoo ◽  
Da-Seul Kim ◽  
Kyoung-Won Ko ◽  
...  
Keyword(s):  
Cell Culture ◽  
Culture Medium ◽  
Density Lipoprotein ◽  
High Yield ◽  
Human Umbilical Cord ◽  
Cell Culture Medium ◽  
Human Coronary Artery ◽  
Isolation Methods ◽  
Regenerative Activity ◽  
Yield And Purity

Exosomes derived from mesenchymal stem cells (MSCs) have been studied as vital components of regenerative medicine. Typically, various isolation methods of exosomes from cell culture medium have been developed to increase the isolation yield of exosomes. Moreover, the exosome-depletion process of serum has been considered to result in clinically active and highly purified exosomes from the cell culture medium. Our aim was to compare isolation methods, ultracentrifuge (UC)-based conventional method, and tangential flow filtration (TFF) system-based method for separation with high yield, and the bioactivity of the exosome according to the purity of MSC-derived exosome was determined by the ratio of Fetal bovine serum (FBS)-derived exosome to MSC-derived exosome depending on exosome depletion processes of FBS. The TFF-based isolation yield of exosome derived from human umbilical cord MSC (UCMSC) increased two orders (92.5 times) compared to UC-based isolation method. Moreover, by optimizing the process of depleting FBS-derived exosome, the purity of UCMSC-derived exosome, evaluated using the expression level of MSC exosome surface marker (CD73), was about 15.6 times enhanced and the concentration of low-density lipoprotein-cholesterol (LDL-c), known as impurities resulting from FBS, proved to be negligibly detected. The wound healing and angiogenic effects of highly purified UCMSC-derived exosomes were improved about 23.1% and 71.4%, respectively, with human coronary artery endothelial cells (HCAEC). It suggests that the defined MSC exosome with high yield and purity could increase regenerative activity.

Three‐dimensional cancer cell culture in high‐yield multiscale scaffolds by shear spinning

2018 ◽  
Vol 35 (2) ◽  
pp. e2750 ◽  
Author(s):  
Ahmed A. Ahmed ◽  
CJ Luo ◽  
Sandra Perez‐Garrido ◽  
Connor R. Browse ◽  
Christopher Thrasivoulou ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cancer Cell ◽  
Three Dimensional ◽  
High Yield ◽  
Shear Spinning

Pre-clinical development of cell culture (Vero)-derived H5N1 pandemic vaccines

2008 ◽  
Vol 389 (5) ◽  
Author(s):  
M. Keith Howard ◽  
Otfried Kistner ◽  
P. Noel Barrett
Keyword(s):  
Cell Culture ◽  
High Yield ◽  
Wild Type Virus ◽  
Effective Vaccine ◽  
Type Virus ◽  
Wild Type ◽  
Yield Production ◽  
Homologous Virus ◽  
Clade 1 ◽  
Influenza H5n1

AbstractThe rapid spread of avian influenza (H5N1) and its transmission to humans has raised the possibility of an imminent pandemic and concerns over the ability of standard influenza vaccine production methods to supply sufficient amounts of an effective vaccine. We report here on a robust and flexible strategy which uses wild-type virus grown in a continuous cell culture (Vero) system to produce an inactivated whole virus vaccine. Candidate vaccines based on clade 1 and clade 2 influenza H5N1 strains, produced at a variety of manufacturing scales, were demonstrated to be highly immunogenic in animal models without the need for adjuvant. The vaccines induce cross-neutralising antibodies and are protective in a mouse challenge model not only against the homologous virus but against other H5N1 strains, including those from other clades. These data indicate that cell culture-grown, whole virus vaccines, based on the wild-type virus, allow the rapid high-yield production of a candidate pandemic vaccine.

The Philips Lecture, 1981 - Targets in biotechnology

1982 ◽  
Vol 214 (1196) ◽  
pp. 289-303 ◽  
Keyword(s):  
Cell Culture ◽  
Recombinant Dna ◽  
Genetic Manipulation ◽  
Animal Cell ◽  
Glucose Isomerase ◽  
High Yield ◽  
Enzyme Specificity ◽  
Blood Proteins ◽  
Bulk Chemicals ◽  
Enhanced Production

Established biotechnologies overcome cost handicaps, particularly aqueous process dilution, through unique advantages. Ancient fermen­tations confer psychopharmacological, palatable or nutritional qualities on drinks and foods. Modern biotechnologies depend on bacterial versa­tility (sewage disposal), enzyme specificity (hydrolases), high value of complex products (antibiotics, vaccines), chirality (amino acids), high yield (citric acid), or, rarely, process intensity (glucose isomerase). Advances in recombinant DNA have already given valuable human proteins. Numerous new targets include hormones, blood proteins and antibodies. Production techniques will include cell fusion and animal cell culture, and new hosts, some eukaryotic, for human genes. Enhanced production of cells, organelles and enzymes will follow, partly to take advantage of the industrial potential of immobilization. Later still, improved engineering and new biological processes should allow competitive production of fine chemicals, protein and other foods, bulk chemicals and fuels. Some products will require simultaneous change and advance in agriculture: interspecific genetic manipulation, plant cell culture and other modern developments will facilitate these.

scholarly journals The Petri Dish-N2B27 Culture Condition Maintains RPE Phenotype by Inhibiting Cell Proliferation and mTOR Activation

2020 ◽  
Vol 2020 ◽  
pp. 1-12
Author(s):  
Hui Lou ◽  
Chunpin Lian ◽  
Fanjun Shi ◽  
Liqun Chen ◽  
Sicheng Qian ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Proliferation ◽  
Growth Medium ◽  
Culture Condition ◽  
Petri Dish ◽  
High Density ◽  
High Yield ◽  
Low Density ◽  
Rpe Cells ◽  
Petri Dishes

Objective. To develop a method for the rapid isolation of rat RPE cells with high yield and maintain its epithelial state in modified culture system. Methods. The eyeballs were incubated with dispase. The retina was isolated with RPE attached and cut into several pieces. Following a brief incubation in growth medium, large RPE sheets can be harvested rapidly. RPE cells were divided into four groups and cultured for several weeks, that is, (1) in cell culture dishes with 10% FBS containing medium (CC dish-FBS), (2) in petri dishes with 10% FBS containing medium (Petri dish-FBS), (3) in cell culture dishes with N2 and B27 containing medium (CC dish-N2B27), and (4) in petri dishes with N2 and B27 containing medium (Petri dish-N2B27). Morphological and biological characteristics were investigated using light microscopy, Q-PCR, and western blot. Results. The retina would curl inwardly during the growth medium incubation period, releasing RPE sheets in the medium. Compared with low density group (5,000 cells/cm2), RPE cells plated at high density (15,000 cells/cm2) can maintain RPE morphology for a more extended period. Meanwhile, plating RPE cells at low density significantly reduced the expression of RPE cell type-specific genes (RPE65, CRALBP, and bestrophin) and increased the expression of EMT-related genes (N-cadherin, fibronectin, and α-SMA), in comparison with the samples from the high density group. The petri dish culture condition reduced cell adhesion and thus inhibited RPE cell proliferation. As compared with other culture conditions, RPE cells in the petri dish-N2B27 condition could maintain RPE phenotype with increased expression of RPE-specific genes and decreased expression of EMT-related genes. The AKT/mTOR pathway was also decreased in petri dish-N2B27 condition. Conclusion. The current study provided an alternative method for easy isolation of RPE cells with high yield and maintenance of its epithelial morphology in the petri dish-N2B27 condition.

scholarly journals Diffusion-Based Separation of Extracellular Vesicles by Nanoporous Membrane Chip

Biosensors ◽  
2021 ◽  
Vol 11 (9) ◽  
pp. 347
Author(s):  
Gijung Kim ◽  
Min Chul Park ◽  
Seonae Jang ◽  
Daeyoung Han ◽  
Hojun Kim ◽  
...  
Keyword(s):  
Cell Culture ◽  
Human Serum ◽  
Extracellular Vesicles ◽  
Culture Media ◽  
High Yield ◽  
Polycarbonate Membrane ◽  
Cell Culture Media ◽  
Nanoporous Membrane ◽  
Selective Filter ◽  
Novel Biomarkers

Extracellular vesicles (EVs) have emerged as novel biomarkers and therapeutic material. However, the small size (~200 nm) of EVs makes efficient separation challenging. Here, a physical/chemical stress-free separation of EVs based on diffusion through a nanoporous membrane chip is presented. A polycarbonate membrane with 200 nm pores, positioned between two chambers, functions as the size-selective filter. Using the chip, EVs from cell culture media and human serum were separated. The separated EVs were analyzed by nanoparticle tracking analysis (NTA), scanning electron microscopy, and immunoblotting. The experimental results proved the selective separation of EVs in cell culture media and human serum. Moreover, the diffusion-based separation showed a high yield of EVs in human serum compared to ultracentrifuge-based separation. The EV recovery rate analyzed from NTA data was 42% for cell culture media samples. We expect the developed method to be a potential tool for EV separation for diagnosis and therapy because it does not require complicated processes such as immune, chemical reaction, and external force and is scalable by increasing the nanoporous membrane size.

scholarly journals A novel three step protocol to isolate extracellular vesicles from plasma or cell culture medium with both high yield and purity

2020 ◽  
Vol 9 (1) ◽  
pp. 1791450 ◽  
Author(s):  
Xiaogang Zhang ◽  
Ellen G. F. Borg ◽  
A. Manuel Liaci ◽  
Harmjan R. Vos ◽  
Willem Stoorvogel
Keyword(s):  
Cell Culture ◽  
Extracellular Vesicles ◽  
Culture Medium ◽  
High Yield ◽  
Cell Culture Medium ◽  
Yield And Purity

Abstract 534: Chemically Defined Cell Culture Media for High Yield Differentiation of Functional Human Cardiac Myocytes

2020 ◽  
Vol 127 (Suppl_1) ◽  
Author(s):  
Davi M Lyra Leite ◽  
Paul W Burridge
Keyword(s):  
Cell Culture ◽  
Recombinant Proteins ◽  
Pluripotent Stem Cell ◽  
Culture Media ◽  
Induced Pluripotent Stem Cell ◽  
High Yield ◽  
Cell Culture Media ◽  
Basal Media ◽  
Induced Pluripotent ◽  
Selection Of

Introduction: Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) are a promising technology for regenerative medicine and pharmacology. However, existing protocols are limited due to the high costs when generating cells at scale, to low yield, and to a lack of protocol replicability. Additionally, the use of animal-derived or recombinant proteins in cell differentiation creates concerns in human compatibility and presents technical challenges due to batch-to-batch variability. Aim: Develop a protocol for functional hiPSC-CM differentiation with high yield without the use of proteins. Methods: We expanded the CDM3 protocol by testing approximately 200 combinations of cell culture media during hiPSC differentiation. The differentiation was induced with a 24h incubation in 6 μM CHIR (d0), followed by 24 h in basal media (d1) and 48h in media supplemented with AA2P and Wnt-C59 (d2-d4). From day 4 onwards, cells were kept in media supplemented with insulin and AA2P, and in some conditions with polyvinyl alcohol. On day 8, hiPSC-CMs were antibiotically selected using geneticin. Tissues were imaged on day 13 to assess myocyte morphology and contraction. Videos were scored from 0 (dead/delaminated cells) to 9 (confluent tissues beating at least 4 times in a 5 sec interval). Results: We identified 6 basal media combinations that yield confluent monolayers on day 13. Out of these, 3 conditions consistently resulted in confluent contractile tissues. Conclusions: Our data demonstrate that a protein-free high yield hiPSC-CM differentiation is possible with the appropriate selection of basal media based on cellular nutritional requirements.

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