Separation of viable from nonviable cells using a discontinuous density gradient

Abstract

When a suspension containing a mixture of viable and nonviable cells is layered over a dense ficoll-metrizoate solution and centrifuged, most of the viable cells are retained at the interface above the dense solution; whereas most of the nonviable cells are distributed in other fractions. The cells recovered from the interface are capable of subsequent growth in culture.

Ultra scale‐down prediction using microwell technology of the industrial scale clarification characteristics by centrifugation of mammalian cell broths

“…Comparison of PSD and centrifugation data is affected by changes in the density difference which can be significant, especially in the transition from viable to non-viable cells and also to debris with each being of higher density (Clynes et al, 1980). A comparison of the fine end of the PSD and the solids remaining in the centrifuge supernatant indicates that for the high V/tS M analyses (Fig.…”

Immunomagnetic bead separation of mononuclear cells from contaminating granulocytes in cryopreserved blood samples

“…Density gradient centrifugation is also used as a method for separation of dead and viable cells [3; 6]. According to our data, isolation of viable cells in cryopreserved leukocytes will result in contamination of viable cell fraction with granulocytes; however, we have found that this procedure can still be effective if granulocytes are removed with magnetic beads before gradient centrifugation (data not shown).…”

Enrichment and Recovery of Mammalian Cells from Contaminated Cultures Using Aqueous Two-Phase Systems

“…1,3 Nonviable cells (i.e., cells present in culture that lack membrane integrity as demonstrated by the inability to exclude dyes), cell fragments, and debris can also be considered contaminants because their presence can bias seeding densities or skew the results of functional assays. 4,5 Cultures that contain a large fraction of nonviable cells (e.g., cultures that are overgrown) may not be usable unless the viable cells can be isolated from the dead cells. 5 Antibiotics, such as penicillin and streptomycin, are used to treat contamination or prevent it prophylactically.…”

“…4,5 Cultures that contain a large fraction of nonviable cells (e.g., cultures that are overgrown) may not be usable unless the viable cells can be isolated from the dead cells. 5 Antibiotics, such as penicillin and streptomycin, are used to treat contamination or prevent it prophylactically. However, these compounds may adversely interfere with normal processes in cultured cells, 6 exacerbate contamination issues by masking poor aseptic technique, or promote antibiotic resistance and the development of resistant microorganisms.…”

“…Typical contaminants include bacteria, yeast, and mold . These microorganisms can affect the way cells behave in culture (e.g., change morphology, growth, and viability) and alter the culture conditions (e.g., through changing pH or by competing for resources in the medium). , Nonviable cells (i.e., cells present in culture that lack membrane integrity as demonstrated by the inability to exclude dyes), cell fragments, and debris can also be considered contaminants because their presence can bias seeding densities or skew the results of functional assays. , Cultures that contain a large fraction of nonviable cells (e.g., cultures that are overgrown) may not be usable unless the viable cells can be isolated from the dead cells …”