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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