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An ultra scale-down analysis of the recovery by dead-end centrifugation of human cells for therapy.

Delahaye M, Lawrence K, Ward SJ, Hoare M - Biotechnol. Bioeng. (2015)

Bottom Line: The cell critical quality attributes studied are the cell membrane integrity and the presence of selected surface markers.Greater hold times and higher RCF values for longer spin times all led to the increased loss of cell membrane integrity.Changes in cell surface markers were significant in some cases but to a lower extent than loss of cell membrane integrity.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemical Engineering, Advanced Centre for Biochemical Engineering, University College London, Torrington Place, London, WC1E 7JE, UK.

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Normalized size distributions (with respect to total cell counts) of OnyCap23 and P4E6; (A) fresh control, (B) hold time (H) = 5 min, centrifugation RCF (R) = 250xg, spin time (S) = 3 min, (C) H = 60 min, R = 10000xg, S = 30 min, (D) H = 120 min, R = 20000xg, S = 30 min. The mean cell diameter, number basis, and the % cell recovery, REC, are given as insets. Temperature = 21°C.
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fig07: Normalized size distributions (with respect to total cell counts) of OnyCap23 and P4E6; (A) fresh control, (B) hold time (H) = 5 min, centrifugation RCF (R) = 250xg, spin time (S) = 3 min, (C) H = 60 min, R = 10000xg, S = 30 min, (D) H = 120 min, R = 20000xg, S = 30 min. The mean cell diameter, number basis, and the % cell recovery, REC, are given as insets. Temperature = 21°C.

Mentions: Image analysis of the total cell population of surface attached cells as a result of centrifugation and resuspension show little change in cell morphology (results not shown here). The size distributions of various cell preparations in suspension were analysed by image analysis with the size related to the diameter of a sphere with equivalent projected area. Sample size distributions for P4E6 cells before centrifugation (Fig. 7) show a monomodal distribution with some clumping. Centrifugation and resuspension appears to just disrupt the clumps resulting in similar size distributions for the various conditions studied ranging from gentle centrifugation (RCF = 250xg for 3 min) of fresh cells to extreme centrifugation (RCF = 20000xg for 30 min) of aged cells. Similar results are noted for OnyCap23 cells except for the appearance of a bimodal distribution at extreme centrifugation conditions of aged cells. Of note is the larger size of OnyCap23 cells with an average cell volume 1.2 fold larger than P4E6.


An ultra scale-down analysis of the recovery by dead-end centrifugation of human cells for therapy.

Delahaye M, Lawrence K, Ward SJ, Hoare M - Biotechnol. Bioeng. (2015)

Normalized size distributions (with respect to total cell counts) of OnyCap23 and P4E6; (A) fresh control, (B) hold time (H) = 5 min, centrifugation RCF (R) = 250xg, spin time (S) = 3 min, (C) H = 60 min, R = 10000xg, S = 30 min, (D) H = 120 min, R = 20000xg, S = 30 min. The mean cell diameter, number basis, and the % cell recovery, REC, are given as insets. Temperature = 21°C.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4402021&req=5

fig07: Normalized size distributions (with respect to total cell counts) of OnyCap23 and P4E6; (A) fresh control, (B) hold time (H) = 5 min, centrifugation RCF (R) = 250xg, spin time (S) = 3 min, (C) H = 60 min, R = 10000xg, S = 30 min, (D) H = 120 min, R = 20000xg, S = 30 min. The mean cell diameter, number basis, and the % cell recovery, REC, are given as insets. Temperature = 21°C.
Mentions: Image analysis of the total cell population of surface attached cells as a result of centrifugation and resuspension show little change in cell morphology (results not shown here). The size distributions of various cell preparations in suspension were analysed by image analysis with the size related to the diameter of a sphere with equivalent projected area. Sample size distributions for P4E6 cells before centrifugation (Fig. 7) show a monomodal distribution with some clumping. Centrifugation and resuspension appears to just disrupt the clumps resulting in similar size distributions for the various conditions studied ranging from gentle centrifugation (RCF = 250xg for 3 min) of fresh cells to extreme centrifugation (RCF = 20000xg for 30 min) of aged cells. Similar results are noted for OnyCap23 cells except for the appearance of a bimodal distribution at extreme centrifugation conditions of aged cells. Of note is the larger size of OnyCap23 cells with an average cell volume 1.2 fold larger than P4E6.

Bottom Line: The cell critical quality attributes studied are the cell membrane integrity and the presence of selected surface markers.Greater hold times and higher RCF values for longer spin times all led to the increased loss of cell membrane integrity.Changes in cell surface markers were significant in some cases but to a lower extent than loss of cell membrane integrity.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemical Engineering, Advanced Centre for Biochemical Engineering, University College London, Torrington Place, London, WC1E 7JE, UK.

Show MeSH
Related in: MedlinePlus