scholarly journals ViableTM AC-2, a New Adult Bovine Serum- and Colostrum-Based Supplement for the Culture of Mammalian Cells

BioTechniques ◽  
1996 ◽  
Vol 20 (4) ◽  
pp. 702-707
Author(s):  
Britta Viander ◽  
Sari Ala-Uotila ◽  
Markku Jalkanen ◽  
Raimo Pakkanen
Keyword(s):  
Cell Line ◽  
Cell Density ◽  
Cell Lines ◽  
Bovine Serum ◽  
Receptor Expression ◽  
Small Scale ◽  
Maximum Cell Density ◽  
Mammary Tumor Cell ◽  
Adult Bovine Serum ◽  
Maximum Cell

In this study we have shown that ViableTM AC-2, a medium based on an ultrafiltrate fraction of bovine colostrum and adult bovine serum, can be used successfully as a fetal bovine serum (FBS) substitute in the culture of several anchorage-dependent and independent cell lines. Of the 15 cell lines cultured in 8% Viable AC-2 in microplates, 10 reached the maximum cell density of 65%-123% of that in 10% FBS, 4 cell lines reached maximum cell density of 35%-65% of that in 10% FBS and only one cell line, a human osteosawoma G-292, grew slowly in Viable AC-2. In a small-scale suspension culture, 8%-15% Viable AC-2 supports the growth of Chinese hamster ovary cells (CHO-K1) on microcarriers in spinner flasks significantly better than 10% FBS. Shionogi mouse mammary tumor cell line (S115) transfected with human α2-adrenergic receptor subtype C2 was used as a model to study recombinant protein production in Viable AC-2-supplemented medium. The results showed that in cell culture flasks and in an ACUSYST-RTM bioreactot, the α2-C2 receptor expression level per mg of total protein was similar in both Viable AC-2 and FBS.

A Comparison of CO2 and N2 Foaming Behaviors of PP in a Visualization System

2020 ◽  
Vol 35 (5) ◽  
pp. 503-517
Author(s):  
Q.-P. Guo ◽  
J. Wang ◽  
C. B. Park
Keyword(s):  
Cell Density ◽  
Nucleating Agent ◽  
Gas Content ◽  
Experimental Conditions ◽  
Maximum Cell Density ◽  
Visualization System ◽  
Cell Nucleation ◽  
Processing Strategies ◽  
Automotive Parts ◽  
Maximum Cell

Abstract Understanding of polypropylene (PP) foaming is critically important to reduce the weight of automotive parts. In this study, we used a batch foaming simulation system with visualization cell, to observe the foaming behaviors of PP that is blown with CO2 and N2 under various experimental conditions. We found that the nucleating agent content, initial temperature, pressure (i. e., gas content), and pressure drop rate during foaming have a significant effect on cell nucleation and cell growth. The cell density and the void fraction of PP foamed with CO2 and N2, respectively, were separately observed and compared. It was found that under the same experimental conditions, the maximum cell density of PP foamed with CO2 was higher than that of PP foamed with N2. However, the maximum cell density of PP foamed with CO2 was determined to be lower than that of PP foamed with N2, when the same gas mole numbers were employed. Based on the experimental results, optimum foaming conditions and effective processing strategies for PP-CO2 system are suggested.

scholarly journals Interleukin 2 high-affinity receptor expression requires two distinct binding proteins.

1987 ◽  
Vol 165 (1) ◽  
pp. 223-238 ◽  
Author(s):  
K Teshigawara ◽  
H M Wang ◽  
K Kato ◽  
K A Smith
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Binding Sites ◽  
Binding Proteins ◽  
Lymphoblastic Leukemia ◽  
Interleukin 2 ◽  
Leukemia Cell ◽  
Receptor Expression ◽  
High Affinity ◽  
Antigen Protein

A cell line established from a patient with acute lymphoblastic leukemia was found to express IL-2 binding sites with a novel, intermediate affinity compared with the characteristic high-affinity IL-2-receptors and low-affinity IL-2 binding sites described previously. Clones were isolated from this cell line that displayed solely this new IL-2-binding protein, and were found to be unreactive with anti-Tac, the mAb that competes with IL-2 for binding. Moreover, these same cloned cells did not express mRNA detectable by hybridization with radiolabeled cDNA encoding the Tac protein. In contrast, the original cell line and similar clones expressed low levels of Tac mRNA and cell surface Tac antigen, both of which could be augmented by exposure to medium conditioned by adult T leukemia cell lines. Particularly noteworthy, induction of Tac antigen expression was paralleled by an increase in the number of high-affinity IL-2-R detectable. Since the expression of the Tac antigen protein by itself makes only for low-affinity IL-2 binding, these data prompted a reevaluation of the structural composition of high-affinity IL-2-R. Analysis of the IL-2-binding proteins expressed by leukemic cell lines lacking high-affinity receptors revealed only a single protein, larger than the Tac antigen protein (Mr = 75,000 vs. 55,000). In contrast, clones induced to express high-affinity receptors had clearly both of these IL-2-binding proteins. Moreover, when IL-2 binding to normal T cells was performed under conditions that favored the proportion of high-affinity receptors occupied, two distinct proteins identical to those already identified on the leukemic cells could be crosslinked covalently to radiolabeled IL-2. The interpretations derived from these varied, assembled data, point to two IL-2-binding proteins, both of which are required for high-affinity IL-2 binding.

scholarly journals Lithocholic bile acid induces apoptosis in human nephroblastoma cells: a non-selective treatment option

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Julian Trah ◽  
Jonas Arand ◽  
Jun Oh ◽  
Laia Pagerols-Raluy ◽  
Magdalena Trochimiuk ◽  
...  
Keyword(s):  
Bile Acid ◽  
Cell Line ◽  
Cell Lines ◽  
Wilms Tumor ◽  
Control Cell ◽  
Kidney Tissue ◽  
Receptor Expression ◽  
Cancer Drug ◽  
Selective Treatment

AbstractLithocholic bile acid (LCA) has been reported to selectively kill cancer cells within many tumor cell lines including neuroblastoma or glioblastoma. Wilms’ tumor shares similarities with neuro- and glioblastoma. Hence, the aim of the study was to evaluate the effects of LCA on nephroblastoma. To test the effects of LCA, nephroblastoma cell line WT CLS1 was used. SK NEP1 was tested as well. It was originally classified as a nephroblastoma cell line but was meanwhile reclassified as an ewing sarcoma cell line. As control cell lines HEK 293 from embryonic kidney and RC 124 from adult kidney tissue as well as podocytes were used. The effects were evaluated using proliferation assay, caspase activity assay, FACS and Western blot. LCA showed a dose and time-dependent selective effect inducing apoptosis in nephroblastoma cells. However, these effects were not limited to the nephroblastoma cell line but also affected control kidney cell lines and the sarcoma cells; only podocytes are significantly less affected by LCA (at dosages < 200 µm). There were no significant differences regarding the TGR5 receptor expression. The study showed that LCA has a strong, yet unselective effect on all used in vitro cell-lines, sparing the highly differentiated podocytes in lower concentrations. Further studies are needed to verify our results before dismissing LCA as an anti-cancer drug.

Patterns of HER-family receptor dimerization in trastuzumab susceptible and trastuzumab resistant cell lines

2007 ◽  
Vol 25 (18_suppl) ◽  
pp. 2533-2533
Author(s):  
R. Dua ◽  
P. Nhonthachit ◽  
C. Rinehart ◽  
C. L. Arteaga ◽  
R. Nahta ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cell Line ◽  
Cell Lines ◽  
Resistant Cell ◽  
Receptor Expression ◽  
Skbr3 Cell ◽  
Bt474 Cell ◽  
Her2 Overexpression ◽  
Trastuzumab Resistance ◽  
Her Family

2533 Background: HER2 overexpression is associated with accelerated disease progression and poor prognosis in breast cancer. Trastuzumab, a monoclonal antibody targeting the extracellular domain of HER2, is effective in the treatment of metastatic breast cancer. However, most patients treated with trastuzumab eventually develop clinical resistance. To investigate the role of HER-family receptors in trastuzumab resistance, we measured HER-family receptor expression, dimerization, and phosphorylation in trastuzumab susceptible and resistant cell lines. Methods: Cell lysates from trastuzumab susceptible and resistant BT474 and SKBR3 cell lines were obtained from the Arteaga and Esteva laboratories. Proximity-based, multiplexed assays were used to detect and quantify HER1, HER2, and HER3 expression and phosphorylation levels, as well as HER1/HER1, HER1/HER2, HER1/HER3, HER2/HER2, and HER2/HER3 dimers. Samples were incubated with a mixture of HER specific antibodies conjugated either with fluorescent reporter tags (eTags), or biotin, which binds a reporter tag releasing agent (chemical scissor). Reporter molecules are released based on proximity to the scissor in a photochemical reaction and separated by capillary gel electrophoresis. Results: In comparison to trastuzumab susceptible parental cell lines, both SKBR3 and BT474 trastuzumab-resistant cell lines displayed upregulated HER1 expression. Resistant BT474 cell lines exhibited markedly increased levels of HER1/HER2 heterodimers. Increases in HER2 phosphorylation in the trastuzumab resistant SKBR3 cell line were observed, consistent with previous studies implicating trastuzumab in the induction of HER2 phosphorylation. Total HER2 and HER3 levels were similar in trastuzumab susceptible and resistant BT474 cell lines. Conclusions: The development of trastuzumab resistance in these cell line models correlated with HER1 expression and the appearance of HER1:HER2 dimers. Since signaling initiated by such heterodimers is ineffectively antagonized by trastuzumab, these data suggest that selection for proliferative signaling mediated by HER1:HER2 dimers may represent a mechanism of trastuzumab resistance in breast cancer. No significant financial relationships to disclose.

EFECTO DE LA ZEOLITA NATURAL CLINOPTILITA SOBRE EL CRECIMIENTO DE DUNALIELLA SALINA (TEODORESCO, 1905) CULTIVADA EN UN FOTOBIORREACTOR DE MÚLTIPLES CÁMARAS OSCILANTES

2013 ◽  
Vol 6 ◽  
Author(s):  
Euler Gallego Cartagena ◽  
Leydis Yohana Herrera Britto ◽  
Lena Judith Manjarrez Rodríguez ◽  
Carmiña Lucía Vargas Zapata
Keyword(s):  
Cell Density ◽  
Dunaliella Salina ◽  
Logarithmic Phase ◽  
Total Chlorophyll ◽  
Maximum Cell Density ◽  
Stages Of Growth ◽  
Maximum Cell ◽  
Level Of Significance ◽  
Duplication Time ◽  
Chloroplastic Pigments

We studied the growth and production of pigment of microalgae Dunaliella salina cultivated in<br />photobioreactor multicamera oscillating outside laboratory conditions to evaluate the effect of different<br />concentrations of zeolite (ZC). Growth was evaluated by cell count and pigment content was performed<br />by spectrophotometric techniques. The results indicate that the concentration of 50 mgL-1 of ZC produced<br />a better stimulus on the growth of the microalga reaching maximum cell density (MDC) of 5.51 ±0.45 x 106<br />celmL-1, growth rate (μ) 0.37 ±0.03 divday-1 and duplication time (Td) of 1.87 ±0.02 days. Likewise,<br />produced a greater increase in the total chlorophyll and carotenoids in the logarithmic phase of values<br />15.554 ±0.77 and 0.50 ±0.01 mgmL-1, respectively. Chloroplastic pigments concentration per volume of<br />culture has a significant correlation with maximum cell density of D. salina treatments based on zeolite at<br />all stages of growth with rMDC, chla.tot= 0.89 y una rMDC, carot.tot=0.926 at a level of significance (p &lt;0.01). The<br />results demonstrated the feasibility of using this product as a suitable substrate for the growth of the<br />microalga, being an innovative alternative and less costly to obtain significant metabolites.

scholarly journals Sulphamoylated estradiol analogue induces antiproliferative activity and apoptosis in breast cell lines

2012 ◽  
Vol 17 (4) ◽  
Author(s):  
Michelle Visagie ◽  
Thandi Mqoco ◽  
Anna Joubert
Keyword(s):  
Estrogen Receptor ◽  
Epithelial Cell ◽  
Cell Line ◽  
Cell Density ◽  
Cell Lines ◽  
Antiproliferative Activity ◽  
Breast Adenocarcinoma ◽  
Epithelial Cell Line ◽  
Antimitotic Activity

AbstractResearch into potential anticancer agents has shown that 2-methoxyestradiol exerts antiproliferative activity in vitro and in vivo in an estrogen receptor-independent manner. Due to its limited biological accessibility and rapid metabolic degradation, several new analogues have been developed in recent years. This study investigated the in vitro effects of a novel in silicodesigned compound (C16) in an estrogen receptor-positive breast adenocarcinoma epithelial cell line (MCF-7), an estrogen receptor-negative breast adenocarcinoma epithelial cell line (MDA-MB-231) and a nontumorigenic breast cell line (MCF-12A). Light microscopy revealed decreased cell density, cells blocked in metaphase and the presence of apoptotic characteristics in all three cell lines after exposure to C16 for 24 h. Polarizationoptical transmitted light differential interference contrast revealed the presence of several rounded cells and decreased cell density. The xCELLigence real-time label-independent approach revealed that C16 exerted antiproliferative activity. Significant inhibition of cell growth was demonstrated after 24 h of exposure to 0.2 μM C16 in all three cell lines. However, the non-tumorigenic MCF-12A cell line recovered extremely well after 48 h when compared to the tumorigenic cell lines. This indicates that C16 acts as an antiproliferative agent, possesses antimitotic activity and induces apoptosis in vitro. These features warrant further investigation.

scholarly journals Comparison of industrial-scale tubular photobioreactor to FRP (fiberglass reinforced plastic) panel photobioreactor on outdoor culture of Nannochloropsis oculata in the marine hatchery

2020 ◽  
Vol 37 (3) ◽  
pp. 303-308
Author(s):  
Yasar Durmaz ◽  
Gökhun Çagatay Erbil
Keyword(s):  
Cell Density ◽  
Marine Fish ◽  
Nannochloropsis Oculata ◽  
Maximum Cell Density ◽  
Tubular Photobioreactor ◽  
Live Feed ◽  
Microalgal Culture ◽  
Reinforced Plastic ◽  
Plastic Panel ◽  
Maximum Cell

Microalgal culture is a key procedure in marine fish hatcheries, but this activity is far from optimized and has several problems remain to be solved. Nannochloropsis oculata are important to live feed organisms, which are used to rear the larvae of marine finfish. N. oculata were cultivated in tubular PBR and FRP panel PBR in a greenhouse. Tubular PBR was reached 701.7 x 106 cells mL-1 as its maximum cell density and FRP panel PBR was reached 245 x 106 cells mL-1 as maximum. Also, estimated maximum dry weights of tubular and FRP panel PBRs were calculated as 3.249 g L-1 and 1.47 g L-1, respectively. Consequently, tubular PBR was showed that it is more efficient than FRP panel PBR in this study.

scholarly journals Isotyping antibodies produced from hybridoma A6G11C9

2018 ◽  
Vol 15 (1) ◽  
pp. 39-44
Author(s):  
Nguyễn Thị Trung ◽  
Trương Nam Hải
Keyword(s):  
Monoclonal Antibodies ◽  
Cell Line ◽  
Cell Density ◽  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Hybrid Cell ◽  
Hybrid Cell Line ◽  
Animal Cells ◽  
Serum Supplement ◽  
Fetal Bovine

Hybridoma technology was discovered in 1975 to produce the monoclonal antibodies. By this way, we can produce a desired antibody in large amounts. From a single hybrid cell line, they grow and develop to produce a monoclonal antibody with large enough quantities to use for the research, treatment and diagnosis. The level of the antibody producing is depended on the cell density and the incubation period. The growth capacity of each hybrid cell line depends on the composition of the substances in the culture medium of animal cells. Among them, fetal bovine serum is the most commonly used serum-supplement for the in vitro cell culture. This is due to it having a very low level of antibodies and containing more growth factors. In the previous studies, the hybrid cell line (designed A6G11C9) was the best one secreting the highest anti-A monoclonal antibodies, whose specificity agglutinaned human red blood cells containing A antigen. This report showed the result of the study on the coditional culture and isotyping of the immunoglobulin. The hybrid cell line A6G11C9 was cultured in the DMEM medium with different level of fetal bovine serum. As the results, this hybridoma line grow in the DMEM medium containing 10% fetal bovine serum is five times faster than in the DMEM medium with 1% fetal bovine serum. The maximum of the cell density are 11.106 cells/ml after 50 hours innoculation. The maximum of the titer from culture supernatant are 1/1024 after 100 hours innoculation. The monoclonal antibodies derived from hybrid cell line A6G11C9 are IgM heavy chain and the kappa light chain.

Transfection of NK Cells with mRNA or Lentivirus Expressing Chimeric Antigen Receptors Results in Highly Efficient Killing of Lymphoid Malignancies and Compares Favorably with Monoclonal Antibody-Directed ADCC.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 1696-1696 ◽  
Author(s):  
Laurent Boissel ◽  
Monica Betancur ◽  
Richard A. Van Etten ◽  
Hans-Georg Klingemann
Keyword(s):  
Monoclonal Antibody ◽  
Nk Cells ◽  
Cell Line ◽  
Cell Lines ◽  
Receptor Expression ◽  
Antigen Receptors ◽  
Chimeric Antigen Receptors ◽  
Highly Efficient ◽  
Lentivirus Transduction ◽  
Anti Cd20

Abstract Abstract 1696 Poster Board I-722 NK cells frequently do not kill malignant lymphoid target cells except through antibody dependent cellular cytotoxicity (ADCC) when combined with monoclonal antibodies (mAb). Here we compared the ability of the human NK cell line NK-92 to lyse lymphoid cell lines and primary chronic lymphoid leukemia (CLL) cells after transfection with either mRNA or lentivirus coding for Chimeric Antigen Receptors (CAR) against CD19 or CD20. Electroporation (Genepulser, Biorad) of 10 μg mRNA (coding for GFP, CD19-CAR or CD20-CAR) into NK-92 cells was performed at 300V and 150 μF (Leuk. Res 2009;33:1255). For lentivirus transduction, CD19-CAR and CD20-CAR were cloned into a pCL20c IRES-GFP vector and lentivirus stocks for GFP, CD19-CAR and CD20-CAR were obtained from 293T packaging cells. NK-92 cells were transduced by two successive rounds of spinfection in the presence of 8 μg/ml protamine sulfate. Mean transfection efficiencies were 57.2% ± 6.6 for electroporation with mRNA, and 32.6% ± 4.1 for lentivirus transduction. The cytotoxity of the transfected/transduced NK-92 cells against the reference Raji cell line was not affected. The NK-92 resistant cell lines SUP-B15 (CD19+/CD20-) and TMD5 (CD19+/CD20+) became highly sensitive to lysis by transfected NK-92 expressing CD19-CAR, while only TMD5 became sensitive to transfected NK-92 expressing CD20-CAR. Importantly, mRNA-transfected NK-92 showed two-fold higher killing compared to lentivirus-transduced NK-92 despite similar receptor expression. As expected, expression of CAR following mRNA transfection was temporary (48 hours), whereas lentivirus transduced NK-92 maintained CAR expression and cytolytic function for indefinite periods in culture. In order to obtain higher target cell killing by lentivirus transduced cells, NK-92 cells could be highly enriched by cell sorting. Subsequently, we compared CAR-directed killing with ADCC on primary B-CLL patient samples, using NK-92 cells genetically modified by lentivirus expressing anti-CD20 CAR versus high affinity Fc receptor-expressing (FcRγIIIA) NK-92 cells combined with the anti-CD20 monoclonal antibody Rituximab. Initially all CLL cells were resistant to killing by NK-92. Anti-CD20 CAR transfected NK-92 cells showed significantly higher cytotoxicity than ADCC against patients' cells, with less patient-to-patient variability. In conclusions, transfection of NK cells with mRNA coding for CAR against lymphoid surface antigens is a highly efficient method that can easily be scaled up to produce clinical grade material. Lentivirus transduced NK-92 cells show stable expression of CAR, but efficient killing requires sorting or selection for transfected cells. CAR-directed killing by NK-92 compares favorably with anti-CD20 mAb mediated ADCC against primary CLL targets and either approach, alone or in combination, could have clinical relevance. Disclosures Klingemann: ZelleRx Corp.: Co-founder and shareholder.

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