Comparisons of microcarrier cell culture processes in one hundred mini-liter spinner flask and fifteen-liter bioreactor cultures

Wu, Suh-Chin; Hsieh, Wen-Chin; Liau, Ming-Yi

doi:10.1007/s004490050544

Cited by 13 publications

(12 citation statements)

References 7 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Wu et al, 10 who investigated Vero cell growth on Cytodex1 microcarriers, found a close correlation between decrease in suspension cell number and increase in cells attaching to microcarriers. This is in opposite to our findings with significantly different kinetic constants k sus and k at under all cultivation conditions.…”

Section: Attachment Behaviormentioning

confidence: 91%

“…The same result was found by Forestell et al 6 for MRC-5 cells. Ng et al, 8 Wu et al, 10 and Hu et al 11 showed that attachment rate on Cytodex1 microcarriers was highest at a pH of 7.1-7.2, whereas attachment to uncharged macroporous Cultisphere microcarriers was unaffected by changes in pH value over a wide range. At the optimum pH, the higher electrostatic forces of the positively charged Cytodex1 microcarriers probably reduced detachment of nonspreading cells from the microcarriers' surface.…”

Section: Introductionmentioning

confidence: 96%

“…In general, the kinetic constant was directly proportional to the initial cell-to-bead ratio. Wu et al 10 found, under constant initial conditions, slower cell attachment for a large-scale bioreactor (15 L) compared with a 100-mL vessel. These studies did not report an increase in the number of dead cells in suspension during the attachment process.…”

Section: Introductionmentioning

confidence: 99%

“…In all studies, [8][9][10][11] the number of viable cells in suspension and the number of cells attached to microcarriers was determined using a method first introduced by Sanford et al 12 and later modified by van Wezel 13 and Levine et al 14 For attached cells on microcarriers, a certain volume of 0.1 M citrate and 0.1% crystal violet was added to a representative sample and the number of released and stained nuclei counted in a haemocytometer. The number of attached cells was corrected using the recorded volume of settled microcarriers in the sample as reported by Napapetian et al 15 and Mukhopadhyay et al 16 Kong et al 17 and Wu et al 10 determined a half-life time for cell attachment that did not differ between the two complementary kinetics of cells attachment and decrease in suspension cells. Both authors concluded that the attachment of cells on microcarriers can be described with first-order kinetics.…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Growth behavior of number distributed adherent MDCK cells for optimization in microcarrier cultures

Böck

Sann

Schulze‐Horsel

et al. 2009

Biotechnology Progress

View full text Add to dashboard Cite

An assay for measuring the number of adherent cells on microcarriers that is independent from dilution errors in sample preparation was used to investigate attachment dynamics and cell growth. It could be shown that the recovery of seeded cells is a function of the specific rates of cell attachment and cell death, and finally a function of the initial cell-to-bead ratio. An unstructured, segregated population balance model was developed that considers individual classes of microcarriers covered by 1-220 cells/bead. The model describes the distribution of initially attached cells and their growth in a microcarrier system. The model distinguishes between subpopulations of dividing and nondividing cells and describes in a detailed way cell attachment, cell growth, density-dependent growth inhibition, and basic metabolism of Madin-Darby canine kidney cells used in influenza vaccine manufacturing. To obtain a model approach that is suitable for process control applications, a reduced growth model without cell subpopulations, but with a formulation of the specific cell growth rate as a function of the initial cell distribution on microcarriers after seeding was developed. With both model approaches, the fraction of growth-inhibited cells could be predicted. Simulation results of two cultivations with a different number of initially seeded cells showed that the growth kinetics of adherent cells at the given cultivation conditions is mainly determined by the range of disparity in the initial distribution of cells on microcarriers after attachment.

show abstract

Section: Attachment Behaviormentioning

confidence: 91%

Section: Introductionmentioning

confidence: 96%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Growth behavior of number distributed adherent MDCK cells for optimization in microcarrier cultures

Böck

Sann

Schulze‐Horsel

et al. 2009

Biotechnology Progress

View full text Add to dashboard Cite

show abstract

“…For this reason, the growing of cells in MCs is widely used to obtain the vaccine viruses such as rabies ( Frazzati-Gallina NM et al 200;Mendonça RZ et al 1993 ), chicken viruses (43), Japanese encephalitis virus ( Wu SC, Huang Gyl. 2000;Wu S-C et al 1998 ), measlesvirus (Mendonça RZ et al 1994 ), varicella virus ( Chun BH, Chung SI. 2001 ), bovine respiratory syncytial virus ( Moran E. 1999 ).…”

Section: Discussionmentioning

confidence: 99%

Parameters for MDBK cell growth on microcarriers and BoHV-1 virus production

Freitas¹,

Mozzer²,

Freitas³

et al. 2017

VR&R

View full text Add to dashboard Cite

Bovine herpes virus 1 (BoHV-1) is an important veterinary agent , which causes infectious bovine rhinotracheitis. This disease affects the respiratory tract or genitals, causing weight loss, reduced milk production and abortion. Several vaccines against BoHV-1 have been developed. In this paper, we study the parameters for MDBK growth on microcarriers (Cytodex 1) and for BoHV-1 virus production. The cell culture attached to microcarriers is an efficient method to enlarge the surface of cell growth and for large-scale cell production. Our studies reveal that MDBK adhered to MCs in 30 minutes and that initial agitation of culture did not influence on the efficiency of adhesion or cell growth. In our experiments, we detected no relevant influence of agitation on initial cell adhesion of MDBK to MCs. The maximum cell yield was similar to all initial conditions of agitation studied. The maximum yield obtained in culture started with 15, 20 and 30 cells / MC, was respectively.8.7 x105, 9.3 x105 and 9.8 x105 cells / ml . The cellular distribution on the MCs at the beginning of the culture was more heterogeneous in higher initial densities. After three medium exchanges during MDBK cell culture, the increase in the final yield was 100% higher than that from culture performed without medium change (0.93 x 106 cells / mL). Replacing 50% of the culture medium with fresh medium after 24 hours of growth, the concentration of glucose (5 mM) and glutamine (1.8 mM) were almost completely restored. In these studies, BoHV-1 infections of MDBK were performed after 48, 72 and 86 hours with daily exchanges of 50% of the medium. The increase in viral titer was proportional to the number of viable cells present at the time of infection. The best result of BoHV-1 production was achieved when the infection was performed from 86 hours of cell culture, reaching about 3.7 x108 (TCID50/ml) after 24-48 hours of infection, being on average four times higher when compared to the culture in which the infection was performed with 48 hours of culture and approximately 2 times greater than the crop whose infection occurred after 72 hours of culture. The best yields are obtained when viral infections were performed on cultures with higher cell densities. The best result for the production of BoHV-1 occurred with 10 MOI, 48 hours after infection, which yield was 24 and 41% superior to the MOI 0.1 and 1, respectively.

show abstract

Monoclonal Antibody Production Using a New Supermacroporous Cryogel Bioreactor

Nilsang¹,

Nandakumar²,

Galaev³

et al. 2007

Biotechnol Progress

View full text Add to dashboard Cite

A supermacroporous cryogel bioreactor has been developed to culture hybridoma cells for long-term continuous production of monoclonal antibodies (mAb). Hybridoma clone M2139, secreting antibodies against J1 epitope (GERGAAGIAGPK; amino acids, 551-564) of collagen type II, are immobilized in the porous bed matrix of a cryogel column (10 mL bed volume). The cells got attached to the matrix within 48 h after inoculation and grew as a confluent sheet inside the cryogel matrix. Cells were in the lag phase for 15 days and secreted mAb into the circulation medium. Glucose consumption and lactic acid production were also monitored, and during the exponential phase (approximately 20 days), the hybridoma cell line consumed 0.75 mM day-1 glucose, produced 2.48 mM day-1 lactic acid, and produced 6.5 microg mL-1 day-1 mAb during the exponential phase. The mAb concentration reached 130 microg mL-1 after continuous run of the cryogel column for 36 days. The yield of the mAb after purification was 67.5 mg L-1, which was three times greater than the mAb yield obtained from T-flask batch cultivation. Even after the exchange of medium reservoir, cells in the cryogel column were still active and had relatively stable mAb production for an extended period of time. The bioreactor was operated continuously for 55 days without any contamination. The results from ELISA as well as arthritis experiments demonstrate that the antibodies secreted by cells grown on the cryogel column did not differ from antibodies purified from the cells grown in commercial CL-1000 culture flasks. Thus, supermacroporous cryogels can be useful as a supporting material for productive hybridoma cell culture. Cells were found to be viable inside the porous matrix of the cryogel during the study period and secreted antibodies continuously. The antibodies thus obtained from the cryogel reactor were found to be functionally active in vivo, as demonstrated by their capacity to induce arthritis in mice.

show abstract

Comparisons of microcarrier cell culture processes in one hundred mini-liter spinner flask and fifteen-liter bioreactor cultures

Cited by 13 publications

References 7 publications

Growth behavior of number distributed adherent MDCK cells for optimization in microcarrier cultures

Growth behavior of number distributed adherent MDCK cells for optimization in microcarrier cultures

Parameters for MDBK cell growth on microcarriers and BoHV-1 virus production

Monoclonal Antibody Production Using a New Supermacroporous Cryogel Bioreactor

Contact Info

Product

Resources

About