scholarly journals 292. Towards Large-Scale Manufacturing of Adeno-Associated Virus by Transient Transfection of HEK293 Suspension Cells in a Stirred Tank Bioreactor Using Serum-Free Medium

2016 ◽  
Vol 24 ◽  
pp. S117-S118 ◽  
Author(s):  
Parminder Chahal ◽  
Érica Alessandra Schulze ◽  
Alice Bernier ◽  
Stephane Lanthier ◽  
Nathalie Coulombe ◽  
...  
Keyword(s):  
Large Scale ◽  
Transient Transfection ◽  
Stirred Tank ◽  
Serum Free Medium ◽  
Free Medium ◽  
Suspension Cells ◽  
Stirred Tank Bioreactor ◽  
Adeno Associated Virus ◽  
Serum Free

Serum free medium for large scale microcarrier culture system of hmsc towards clinical cell-based therapy

Cytotherapy ◽  
2020 ◽  
Vol 22 (5) ◽  
pp. S79
Author(s):  
D. Fiorentini ◽  
S. Daniliuc ◽  
M. Genser-Nir ◽  
Y. Miropolski ◽  
M. Sharovetsky ◽  
...  
Keyword(s):  
Culture System ◽  
Large Scale ◽  
Microcarrier Culture ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Cell Based Therapy

scholarly journals Establishment of a Suspension MDBK Cell Line in Serum-Free Medium for Production of Bovine Alphaherpesvirus-1

Vaccines ◽  
2021 ◽  
Vol 9 (9) ◽  
pp. 1006
Author(s):  
Pengpeng Wang ◽  
Shulin Huang ◽  
Chengwu Hao ◽  
Zhanhui Wang ◽  
Haoran Zhao ◽  
...  
Keyword(s):  
Cell Line ◽  
Neutralizing Antibodies ◽  
Scale Up ◽  
Virus Titer ◽  
Mdbk Cell ◽  
Serum Free Medium ◽  
Free Medium ◽  
Suspension Cells ◽  
Serum Free ◽  
Mdbk Cells

The Madin–Darby bovine kidney (MDBK) cell line is currently used for the production of bovine alphaherpesvirus-1 (BoHV-1) vaccine. For the purpose of vaccine manufacturing, suspension cells are preferred over adherent ones due to simplified sub-cultivation and an easier scale-up process, both of which could significantly reduce production cost. This study aimed to establish a procedure for the culture of BoHV-1 in the suspended MDBK cell line in serum-free medium. We screened several commercially available serum-free media and chose ST503 for subsequent experiments. We successfully adapted the adherent MDBK cells to suspended growth in ST503 in the absence of serum. The maximum density of suspension-adapted MDBK cells could reach 2.5 × 107 cells/mL in ST503 medium with optimal conditions. The average size of suspension-adapted cells increased to 18 ± 1 µm from 16 ± 1 µm. Moreover, we examined tumorigenicity of the suspended cells and found no sign of tumorigenicity post adaptation. Next, we developed a protocol for the culture of BoHV-1 in the cell line described above and found that ultrasonic treatment could facilitate virus release and enhance virus yield by 11-fold, with the virus titer reaching 8.0 ± 0.2 log10TCID50/mL. Most importantly, the prototype inactivated BoHV-1 vaccine we generated using the suspension cultures of MDBK cells induced neutralizing antibodies to a titer comparable to that of the commercial inactivated BoHV-1 vaccine. Overall, we established and optimized a protocol for the production of inactivated BoHV-1 vaccine in MDBK cells adapted for suspension culture, which provides insights for future large-scale manufacturing of BoHV-1 vaccine.

Selective generation of antigen-specific human hybridomas optimized for large scale growth in serum-free medium

1992 ◽  
Vol 154 (1) ◽  
pp. 21-26 ◽  
Author(s):  
Alois B. Lang ◽  
Ulrich Schürch ◽  
Frank Zimmermann ◽  
Urs Bruderer
Keyword(s):  
Large Scale ◽  
Serum Free Medium ◽  
Free Medium ◽  
Scale Growth ◽  
Serum Free ◽  
Selective Generation

Increasing Vero viable cell densities for yellow fever virus production in stirred-tank bioreactors using serum-free medium

Vaccine ◽  
2015 ◽  
Vol 33 (35) ◽  
pp. 4288-4291 ◽  
Author(s):  
Diogo A. Mattos ◽  
Marlon V. Silva ◽  
Luciane P. Gaspar ◽  
Leda R. Castilho
Keyword(s):  
Yellow Fever ◽  
Yellow Fever Virus ◽  
Virus Production ◽  
Viable Cell ◽  
Fever Virus ◽  
Stirred Tank ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Cell Densities

Exogeneous factors are not necessary in attachment, spreading and polarization of hela cells

1978 ◽  
Vol 36 (2) ◽  
pp. 310-311
Author(s):  
W. Liebrich
Keyword(s):  
Hela Cells ◽  
Calf Serum ◽  
Glass Tube ◽  
Serum Free Medium ◽  
Nacl Solution ◽  
Free Medium ◽  
Minimum Essential Medium ◽  
Serum Free ◽  
All Solutions ◽  
Glass Support

HeLa cells were grown for 2-3 days in EAGLE'S minimum essential medium with 10% calf serum (S-MEM; Seromed, München) and then incubated for 24 hours in serum free medium (MEM). After detaching the cells with a solution of 0. 14 % EDTA and 0. 07 % trypsin (Difco, 1 : 250) they were suspended in various solutions (S-MEM = control, MEM, buffered salt solutions with or without Me++ions, 0. 9 % NaCl solution) and allowed to settle on glass tube slips (Leighton-tubes). After 5, 10, 15, 20, 25, 30, 1 45, 60 minutes 2, 3, 4, 5 hours cells were prepared for scanning electron microscopy as described by Paweletz and Schroeter. The preparations were examined in a Jeol SEM (JSM-U3) at 25 KV without tilting.The suspended spherical HeLa cells are able to adhere to the glass support in all solutions. The rate of attachment, however, is faster in solutions without serum than in the control. The latter is in agreement with the findings of other authors.

Hemoglobin Enhances the Binding of Bacterial Endotoxin to Human Endothelial Cells

1996 ◽  
Vol 76 (02) ◽  
pp. 258-262 ◽  
Author(s):  
Robert I Roth
Keyword(s):  
Endothelial Cells ◽  
Binding Protein ◽  
Bacterial Endotoxin ◽  
Dependent Manner ◽  
Serum Free Medium ◽  
Free Medium ◽  
Human Endothelial Cells ◽  
Serum Factors ◽  
Serum Free ◽  
Lps Binding

SummaryHuman endothelial cells, when incubated with bacterial endotoxin (lipopolysaccharide, LPS), modify their surface in association with prominent production of procoagulant tissue factor (TF) activity. This deleterious biological effect of LPS has been shown previously to be enhanced approximately 10-fold by the presence of hemoglobin (Hb), a recently recognized LPS binding protein that causes disaggregation of LPS and increases the biological activity of LPS in a number of in vitro assays. The present study was performed to test the hypothesis that Hb enhances the LPS-induced procoagulant activity of human umbilical vein endothelial cells (HUVEC) by increasing LPS binding to the cells. The binding of 3H-LPS to HUVEC was determined in the absence or presence of Hb or two other known LPS-binding proteins, human serum albumin (HSA) and IgG. LPS binding was substantially increased in the presence of Hb, in a Hb concentration-dependent manner, but was not increased by HSA or IgG. Hb enhancement of LPS binding was observed in serum-free medium, indicating that there was no additional requirement for any of the serum factors known to participate in the interaction of LPS with cells (e.g., lipopolysaccharide (LPS)-binding protein (LBP) and soluble CD14 (sCD14)). Hb enhancement of LPS binding also was observed in the more physiologic condition of 100% plasma. LPS-induced TF activity was stimulated by Hb, but not by HSA or IgG. In serum-free medium, TF activity was not stimulated under any of the conditions tested. Ultrafiltration of LPS was dramatically increased after incubation with Hb but not with HSA or IgG, suggesting that LPS disaggregation by Hb was responsible for the enhanced binding of LPS to HUVEC and the subsequent stimulation of TF activity.

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