Potential and problems of animal cells in suspension culture

Leist, C.; Meyer, Hans‐Peter; Fiechter, Armin

doi:10.1016/0168-1656(90)90049-h

Cited by 34 publications

(10 citation statements)

References 126 publications

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“…Serum is routinely added to the cryopreservation media as a source of nutrients, and other undefi ned factors, in spite of scientifi c and technical disadvantages to its inclusion, its high cost, and the increasing availability of serum-free alternatives [17][18][19][20]. Obviously, for in vivo clinical use, the elimination of animal serum proteins and all possible sources of infectious diseases such as hepatitis, human immunodefi ciency virus (HIV), and bovine spongiform encephalopathy (BSE) is a prerequisite [21][22][23].…”

Section: Introductionmentioning

confidence: 99%

Evaluation of Methylcellulose and Dimethyl Sulfoxide as the Cryoprotectants in a Serum-Free Freezing Media for Cryopreservation of Adipose-Derived Adult Stem Cells

Thirumala

Gimble

Devireddy

2010

Stem Cells and Development

102

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Developing effective techniques for the cryopreservation of human adipose-derived adult stem cells (ASCs) could increase the usefulness of these cells in tissue engineering and regenerative medicine. To this end, we investigated the post-freeze/thaw viability and apoptotic behavior of Passage 1 (P1) adult stem cells (ASCs) in 11 different media: (i) the traditional media containing Dulbecco's modifi ed Eagle's medium (DMEM) with 80% fetal calf serum (FCS) and 10% dimethyl sulfoxide (DMSO), (ii) DMEM with 80% human serum (HS) and 10% DMSO, (iii) DMEM with 1% methyl cellulose (MC) and 10% of either HS or FCS or DMSO, and (iv) DMEM with 0%, 2%, 4%, 6%, 8%, or 10% DMSO. Approximately 1 mL (10 6 cells/mL) of P1 ASCs were frozen overnight in a −80°C freezer and stored in liquid nitrogen for 2 weeks before being rapidly thawed in a 37°C water bath (1-2 min of agitation), resuspended in culture media, and seeded in separate wells of a 6-well plate for a 24-h incubation period at 37°C. After 24 h, the thawed samples were analyzed by bright-fi eld microscopy and fl ow cytometry. The results suggest that the absence of DMSO (and the presence of MC) signifi cantly increases the fraction of apoptotic and/or necrotic ASCs. However, the percentage of viable cells obtained with 2% DMSO and DMEM was comparable with that obtained in freezing media with 10% DMSO and 80% serum (HS or FCS), that is, ~84% ± 5% and ~84% ± 8%, respectively. Adipogenic and osteogenic differentiation behavior of the frozen thawed cells was also assessed using histochemical staining. Our results suggest that post-thaw ASC viability, adipogenic and osteogenic differentiability can be maintained even when they are frozen in the absence of serum but with a minimal concentration of 2% DMSO in DMEM.

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Section: Introductionmentioning

confidence: 99%

Evaluation of Methylcellulose and Dimethyl Sulfoxide as the Cryoprotectants in a Serum-Free Freezing Media for Cryopreservation of Adipose-Derived Adult Stem Cells

Thirumala

Gimble

Devireddy

2010

Stem Cells and Development

102

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“…MSCs encapsulated in 3‐D porous microcarrier beads are protected from shear stresses, although diffusion of oxygen and nutrients within beads can be limited. Furthermore, one challenge associated with microcarrier culture is optimization of cell‐harvesting protocols to guarantee efficient cell‐bead separation and high cell recovery without compromising viability or functionality . (3) MSCs cultured on 3‐D matrices fabricated using polymers, ceramic materials, or decellularized matrices .…”

Section: Introductionmentioning

confidence: 99%

Three‐dimensional carbon nanotube scaffolds for long‐term maintenance and expansion of human mesenchymal stem cells

Lalwani

D’Agati

Gopalan

et al. 2017

J Biomedical Materials Res

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Expansion of mesenchymal stem cells (MSCs) and maintenance of their self-renewal capacity in vitro requires specialized robust cell culture systems. Conventional approaches using animal-derived or artificial matrices and a cocktail of growth factors have limitations such as consistency, scalability, pathogenicity, and loss of MSC phenotype. Herein, we report the use of all-carbon 3-D single- and multiwalled carbon nanotube scaffolds (SWCNTs and MWCNTs) as artificial matrices for long-term maintenance and expansion of human MSCs. Three-dimensional SWCNT and MWCNT scaffolds were fabricated using a novel radical initiated thermal cross-linking method that covalently cross-links CNTs to form 3-D macroporous all-carbon architectures. Adipose-derived human MSCs showed good cell viability, attachment, proliferation, and infiltration in MWCNT and SWCNT scaffolds comparable to poly(lactic-co-glycolic) acid (PLGA) scaffolds (baseline control). ADSCs retained stem cell phenotype after 30 days and satisfied the International Society for Cellular Therapy's (ISCT) minimal criteria for MSCs. Post expansion, (1) ADSCs showed in vitro adherence to tissue culture polystyrene (TCPS); (2) MSC surface antigen expression [CD14(-), CD19(-), CD34(-), CD45(-), CD73(+), CD90(+), CD105(+)]; and (3) trilineage differentiation into osteoblasts, adipocytes, and chondrocytes. Results show that cross-linked 3-D MWCNTs and SWCNTs scaffolds are suitable for ex vivo expansion and maintenance of MSCs for therapeutic applications. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1927-1939, 2017.

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“…Packed‐bed and hollow fiber bioreactors have been commonly used for perfusion culture of immobilized mammalian cells 7. High cell densities of 10 7 –10 8 cells per milliliter can be easily achieved in immobilized cell cultures, extending the productive lifetime of cells and resulting in increased mAb productivity 8, 9. Disadvantages of perfusion culture include their complex nature and difficulty in scale up.…”

Section: Introductionmentioning

confidence: 99%

Supermacroporous polymer‐based cryogel bioreactor for monoclonal antibody production in continuous culture using hybridoma cells

Jain

Karande

Kumar

2010

Biotechnology Progress

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Cryogel matrices composed of different polymeric blends were synthesized, yielding a unique combination of hydrophilicity and hydrophobicity with the presence or absence of charged surface. Four such cryogel matrices composed of polyacrylamide-chitosan (PAAC), poly(N-isopropylacrylamide)-chitosan, polyacrylonitrile (PAN), and poly(N-isopropylacrylamide) were tested for growth of different hybridoma cell lines and production of antibody in static culture. All the matrices were capable for the adherence of hybridoma cell lines 6A4D7, B7B10, and H9E10 to the polymeric surfaces as well as for the efficient monoclonal antibody (mAb) production. PAAC proved to be relatively better in terms of both mAb production and cell growth. Further, PAAC cryogel was designed into three different formats, monolith, disks, and beads, and used as packing material for packed-bed bioreactor. Long-term cultivation of 6A4D7 cell line on PAAC cryogel scaffold in all the three formats could be successfully done for a period of 6 weeks under static conditions. Continuous packed-bed bioreactor was setup using 6A4D7 hybridoma cell line in the three reactor formats. The reactors ran continuously for a period of 60 days during which mAb production and metabolism of cells in the bioreactors were monitored periodically. The monolith bioreactor performed most efficiently over a period of 60 days and produced a total of 57.5 mg of antibody in the first 30 days (in 500 mL) with a highest concentration of 115 μg mL(-1) , which is fourfold higher than t-flask culture. The results demonstrate that appropriate chemistry and geometry of the bioreactor matrix for cell growth and immobilization can enhance the reactor productivity.

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Potential and problems of animal cells in suspension culture

Cited by 34 publications

References 126 publications

Evaluation of Methylcellulose and Dimethyl Sulfoxide as the Cryoprotectants in a Serum-Free Freezing Media for Cryopreservation of Adipose-Derived Adult Stem Cells

Evaluation of Methylcellulose and Dimethyl Sulfoxide as the Cryoprotectants in a Serum-Free Freezing Media for Cryopreservation of Adipose-Derived Adult Stem Cells

Three‐dimensional carbon nanotube scaffolds for long‐term maintenance and expansion of human mesenchymal stem cells

Supermacroporous polymer‐based cryogel bioreactor for monoclonal antibody production in continuous culture using hybridoma cells

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