Centrifugation Application Notes
Table 2. Cell Yield and Viability of Different Cell Types and Culturing Stages
Comparison of Conical Tube and Cell Culture Flask Centrifugation
Cell Yield
Cell Viability
Centrifugation Parameters 2 — RPM (x g ) / Time (min)
Culture Stages 3
Cell Culture Flask
Cell Type
Cell Culture Flask 1
Cell Culture Flask
Tube
Tube
HeLa Adherent 10 mL culture
78% 100% 95%
T-75
1200 (329) / 10
S P H H H
1.6E6 / 5 mL 9.8E6 / 5 mL 2.1E6 / 5 mL 1.2E8 / 5 mL 1.0E8 / 5mL
1.2E6 / 5 mL 1.0E6 / 5 mL 2.0E6 / 5 mL 1.0E8 / 5 mL 9.8E7 / 5 mL
87% 99% 95% ND 4 ND
1200 (335) / 10
ND ND
T-25
1200 (329) / 10 1200 (335) / 10
P H H
2.1E6 / mL 2.9E7 / mL 2.6E7 / mL
2.3E6 / mL 2.0E7 / mL 2.0E7 / mL
98% ND ND
99% ND ND
Jurkat Suspension 10 mL culture
T-75
1200 (329) / 10
S P P P H H
2.5E6 / 5 mL 6.0E6 / 5 mL 3.1E6 / 5 mL 10.6E6 / 5 mL 3.0E7 / 5 mL 3.0E7 / 5 mL
1.8E6 / 5 mL 5.3E6 / 5 mL 2.9E6 / 5 mL 8.3E6 / 5 mL 3.1E7 / 5 mL 3.1E7 / 5 mL
30% 96% 96% 91%
20% 94% 92% 88%
1700 (660) / 15 1200 (335) / 10
ND ND
ND ND
T-25
1200 (329) / 10 1200 (335) / 10
P H H
2.1E6 / mL 7.6E6 / mL 7.5E6 / mL
1.5E6 / mL 7.7E6 / mL 6.7E6 / mL
98% ND ND
98% ND ND
Mononuclear Cell Prep from umbilical cord blood
T-75 No Treatment
ND 92% PT 6
97% ND
ND 98%
1200–1500 (266–416) / 10 1200–1500 (329–514) / 10
P P
84% PT 6 ND
80% PT 6 ND
ND 88% PT 6
98% ND
ND 95%
PHA 5 Stimulated
1200–1500 (266–416) / 10 1200–1500 (329–514) / 10
P P
Monocyte (mononuclear cell) from umbilical cord blood, adherent, 10 mL culture Lymphocyte (mononuclear cell) from umbilical cord blood, 16 mL suspension
ND ND ND ND
T-75
1500 (416) / 5
P H P H
ND ND
ND ND ND ND
96% PT 7 97% PT 7
1500 (514) / 5
ND ND
99% PT 7 93% PT 7
T-75 No Treatment
ND 8% PT 6
ND ND
ND ND
1200–1500 (266–416) / 10 1200–1500 (329–514) / 10
P P
16% PT 6 ND
ND 12% PT 6
20% PT 6 ND
ND ND
ND ND
PHA Stimulated
1200–1500 (266–416) / 10 1200–1500 (329–514) / 10
P P
1. T-75: Corning 75 cm 2 canted-neck cell culture flasks; T-25: Corning 25 cm 2 canted-neck cell culture flasks were used for the experiments. 2. All centrifugation runs conducted in room temperature (18º C–25º C). Centrifugation runs with the conical tubes and cell culture flasks were conducted at the same speed and for the same duration of time. 3. Stages: S (Release from Frozen S torage); P ( P assaging, Reseeding, Expansion); H ( H arvesting). 4. ND: No Data. 5. PHA: Phytohemagglutinin, a mitogen for T-cells. 6. % cell PT = 100% - [(cell in supernatant) / (total cell) x 100]. 7. % cell PT = (cell in pellet) / (total cell) x 100.
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