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Results
Attachment and Distribution. After 2 to 4 hours, both Vero cells (Figure 1) and hMSC attached to the Star-Plus
microcarriers in an evenly-distributed fashion across the microcarrier population. No differences were observed
between microcarriers sterilized by autoclaving or gamma-irradiation (Figure 1). Microscopic observation indicated
that virtually all of the cells were attached to >90% of the microcarriers in a very evenly distributed fashion for
both autoclaved and irradiated conditions.
Figure 1
Vero Cell Attachment and Distribution. Evenly-distributed Vero cells are visualized by DAPI-stained nuclei on
Star-Plus microcarriers.
Irradiated Autoclaved
2
Cell Growth. After four days, cell harvest numbers for the Vero cultures reached 1.5 x 105 cells/cm2 with
a doubling time of 32.7 hours in the autoclaved condition and 1.6 x 105 cells/cm2 with a doubling time of
31.0 hours in the irradiated condition (Figures 2, 3). Both conditions resulted in greater than 97% viable cells.
Figure 2
Vero Cell Growth and Doubling Times. By day 4, healthy and uniformly-distributed cell layers formed on
microcarriers in spinner culture under both conditions. Mean values of cells/cm2 +/- SD are shown (n=3).
1.80E+05
1.60E+05
1.40E+05
1.20E+05
cells/cm2
1.00E+05
8.00E+04 Autoclaved
6.00E+04 Irradiated
4.00E+04
2.00E+04
0.00E+00
0 2 4
Days
Cells/cm2
Condition Day 0 Day 2 Day 4 Doubling Time at Day 4, hrs
Autoclaved 2 x 10
4
4.6 x 10
4
1.5 x 10 32.7
5
Figure 3
Vero cell growth on microcarriers. Representative phase contrast images of Vero cells on Star-Plus microcarriers
4 days post cell seeding demonstrate that healthy, uniformly distributed cell layers are present in both conditions
By day 7, cell harvest numbers for hMSCs reached 7 x 104 cells/cm2 and 8 x 104 cells/cm2 with doubling times
of 37 and 42 hours for the FBS-containing and xeno-free media conditions, respectively (Figure 4). A very
even distribution of cells across the microcarrier population was observed. Bridging of cells and microcarriers
is observed later in the culture period, allowing for cells to grow in the 3-dimensional (3D) space between
microcarriers (Figure 5). Both conditions achieved greater than 95% viable cells. The hMSCs retained their
differentiation capacity after growth and harvest from the Star-Plus microcarriers. Standard adipogenic and
osteogenic differentiation assays were performed. Cells retained their ability to differentiate into adipocytes
and osteocytes when grown on Star-Plus microcarriers (Figure 6).
www.pall.com/biopharm 3
Figure 4
Growth and harvest of hMSCs in stirred-tank bioreactors. Arrows indicate when medium exchanges
(80% of total volume) occurred.
FBS-Containing Medium Xeno-Free Medium
8 0.8 9 0.9
7 0.7 8 0.8
Figure 5
hMSC Attachment and Distribution. Fluorescent images of hMSC DAPI-stained nuclei on Star-Plus microcarriers
at day 7 (100x magnification).
Figure 6
hMSCs grown in bioreactors retain differentiation capacity. hMSCs grown on Star-Plus microcarriers
in serum-containing medium (A) and xeno-free medium (B) were harvested, plated onto flatware and
exposed to differentiation medium.
A.
FBS-Containing Medium B. Xeno-Free Medium
Adipocytes Osteocytes Adipocytes Osteocytes
Control
Diff.
Medium
4
Conclusion
Star-Plus microcarriers were not only designed to be animal component-free, but also to allow for excellent
attachment and growth characteristics from a number of cell line and media combinations. An additional
requirement of the design was that the microcarrier be irradiation-tolerant. These attributes enable greater
flexibility in how the microcarriers are incorporated into different work flows and processes. Results herein
show that attachment, distribution and growth of Vero cells in spinner cultures grown on irradiated Star-Plus
microcarriers were similar to that of the autoclaved control. hMSCs grown on Star-Plus microcarriers in both
FBS-containing and xeno-free media exhibited uniform attachment and distribution across the microcarrier
population, resulting in excellent growth and viability, and importantly retained their differentiation capacity
post-harvest from the microcarriers. Pall Life Sciences new Star-Plus charged microcarriers are excellent
substrates for such applications as cell expansion for the production of cell-based therapies and vaccine
manufacturing.
The information provided in this literature was reviewed for accuracy at the time of publication. Product data may
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