GE Healthcare
Cell culture media
Xuri™ Growth Factors
Growth factors stimulate cellular proliferation and are widely
used for the expansion of T lymphocytes in cell therapy and
research applications. Xuri Growth Factors are produced
under GMP conditions and enable the rapid development of
stable and reproducible cell expansion protocols.
Rapid process development
Each lot of Xuri Growth Factor includes clear information on
biological activity to improve your control over cultivation
variability. This information enables you to standardize your
process and deliver reproducible cell expansion without the
need for revalidation of each new lot.
High quality
Xuri Growth Factors are produced under a certified GMP
license that is regularly qualified by the licensing authorities.
The manufacturing process is in accordance to International
Conference on Harmonization (ICH) guidelines and the release
procedure is executed under a quality management system
following ISO13485. With an endotoxin level of < 25 EU/mg,
the safety of the cells used for treating the patients are the
declared focus of Xuri Growth Factors.
USP <1043> compliance
A smooth regulatory submission process is key when dealing
with cell therapy requirements. To help users assess and
document their production processes, a comprehensive
documentation support file that follows USP <1043> requirements
on ancillary material for cell, gene and tissue-engineered
products is provided (1).
Xuri IL-2
Xuri IL-2 is a reliable solution for the expansion of T lymphocytes
(Fig 2). Xuri IL-2 includes information on biological activity,
which improves reproducibility and removes the need for
IL-2 revalidation.
Data file, 29108247 AB
Fig 1. Xuri IL-2 and IL-2 provide reliable activation of T lymphocytes for
efficient cell expansion.
Xuri IL-2 offers:
•
•
•
•
•
Minimized process development time
Improved scale-out capacity
Reproducible performance without revalidation of each lot
Established protocol in Xuri Cell Expansion System
Dedicated cell therapy ancillary product following
USP <1043> for ex vivo cultivation
Xuri IL-2 supports compliance with USP<1043>(1). A comprehensive
product documentation support file is available to facilitate risk
assessment and validation for use in cell therapy manufacturing.
To further accelerate development of the cell expansion
process, a detailed, straight-forward protocol for the
expansion of T cells using Xuri IL-2 and the Xuri Cell Expansion
System W5 is provided. This protocol delivers reproducible
cell proliferation results in static and perfusion cultures (Fig 2).
Even after 14 d of culture, the majority of cells were in the
early/intermediate stages of differentiation based on CD27
and CD28 expression patterns (Fig 3) without detectable
senescence of the cells.
Xuri Growth Factor products meet USP <1043> “ancillary materials for cell, gene, and
tissue-engineered products”, within the responsibilities applicable to a supplier. Other
aspects of USP <1043> will be the responsibility of the end-user to assess. GE Healthcare
cannot fulfil USP <1043> in regards to application and therapy specific aspects (e.g., use
in finished therapeutic, assessment of removal from a finished therapeutic and possibly
biocompatibility, cytotoxicity or adventitious agent testing).
1
Perfusion culture (Xuri W5)
2.5 × 1010
Seeding density
For static culture at 2.26 × 108 cells
2.0 × 1010
For Xuri W5 at 5 × 108 cells
2.0 × 109
1.5 × 1010
Static culture (T225)
1.5 × 109
Day 0
1.0 × 109
Day 10
Day 0
6.61%
92.7%
Day
Day 0
0
6.61%
CD28
0
92.7%
Day
Day 0
0
6.61%
6.61%
92.7%
92.7%
Day 14
9
5 × 10
28.2%
Day 10 +
CD3+/CD483.6%
15.8%
Day
Day 10
10
15.8%
15.8%
Day
Day 10
10 83.6%
83.6%
Day
Day 14
14 71.2%
28.2%
28.2%
Day
Day 14
14 71.2%
71.2%
28.2%
28.2%
Day 14 71.2%
71.2%
0.400%
28.2%
0.408%
Day 14 71.2%
0.529%
28.2%
0.408%
0
0
Day 10 14
15.8%
83.6%
Day 9 Day 10DayDay
11 Day 1215.8%
Day 13Day
83.6%
6.61%
92.7%
6.61%
92.7%
4
5 Day 0 6
7 15.8% 8 Day 10 9 83.6%
CD28
CD27
15.8%
0.197%
0.197%
0.197%
Day
0
CD27
.1%
95
0.447%0.447%
.0%
95
0.447%
92.7%
0.019%
0.019%
0.197%
.7%
95
0.019%
6.61%
.1%
97
92.7%
.9%
95
.3%
94
CD28
CD28 CD28
CD28
0.447%
0.019%
6.61%
Day 14 71.2%
28.2%
83.6%
CD28
Day 8
3
.8%
95
.3%
93
Viability
Day 7
2
CD28
Day 6
1
Day 3 Day 4 Day 5
15.8%
.3%
94
5.0 × 108
1.0 × 1010
CD3+/CD4+
Typical cell dose per patient (80 kg)
83.6%
0.400%
0.400%
0.400%
Day 10
CD3+/CD8+
0.408%
0.408%
71.2%
0.529%
0.529%
Day 14 0.529%
(C)
CD28
(A)
CD28
CD28
CD28 CD28
CD28
CD28
CD28
0.447%
0.197%
0.400%
0.019%
0.447% flasks) with 5%
0.197%
0.400%plus 200
0.019%
Fig 2. T lymphocytes isolated from peripheral blood were initially cultivated in static cell culture
(T225
human
serum
IU/mlDay
Xuri
IL-2
0.408%
0.529%
0.408%
Day
0
Day 10
14 0.529%
73.3%
CD27
13.3%
CD27
13.9%
77.3% passed
(left graph). Cells were activated using antibody coated beads. Cells were counted from day
three
onwards,
and
expanded
until
they
the number
0.447% 82.0%
0.197%
0.400%
3.64%
0.019%
0.408%
0.529%
Day
0
Day
10
Day
14
8
Day
0
Day
10
Day
14
73.3%
CD27Cell Expansion13.9%
13.3% 9 d under
needed for seeding in a 2 l Cellbag™ bioreactor (5 × 10 ). Cells were transferred and cultivated on a Xuri
System
W5 for
an additional
CD27
CD3+/CD8+
77.3%
0.447% 82.0%
0.197%
0.400%
3.64%
0.019%
0.408%
0.529%
Day
0
10
Day
perfusion. Media composition and Xuri IL-2 concentrations were kept as in static culture (see application
note 28-9650-52Day
available
at www.gelifesciences.
Day
0
Day
10
Day 14
14 73.3%
13.3%
73.3%
CD27
13.3%
13.9%
77.3%
13.9%
77.3%
3.64%
82.0%
3.64%
82.0%
com/xuri). Cell count and viability analysis were performed each day.
CD3+/CD4+
Day 10
CD28
CD28 CD28
CD28 CD28
CD28 CD28
CD28
CD3
CD3
CD3
0.447%
92.7%
15.8%
0.197%
83.6%
0.400%
0.197%
0.197%
0.400%
0.400%
0.019%
0.019%
0.447%
0.019%
0.019%
CD27
0.447%
0.447%
0.197%
0.197%
CD27
0.447%
13.9%
0.197%
CD27
0.447%
13.9%
0.197%
0.019%
3.64%
3.64%
3.64%
CD27
Day
0
Day
0 82.0%
CD27
Day
0
Day
0 82.0%
CD27
Day
0
3.64%
3.64%
Day 0 82.0%
3.37%
3.64%
9.59%
82.0%
3.37%
3.64%
3.37%
3.37%
3.37%
3.37%
3.37%
3.37%
13.9%
13.9%
82.0%
Day
0 82.0%
CD27
13.9%
13.9%
82.0%
CD27
Day
0
CD27
13.9%
1.31%
9.59%
82.0%
13.9%
1.31%
9.59%
9.59%
1.31%
1.31%
9.59%
9.59%
CD27
Day
0
CD27
5.67%
Day
0
CD27
Day
0
CD27
Day
0
CD27
Day
0
1.31%
1.31%
9.59%
1.31%
5.67%
9.59%
1.31%
5.67%
5.67%
28.2%
0.408%
71.2%
0.529%
0.408%
0.408%
0.529%
0.529%
Day 10
CD3+/CD8+
0.400%
0.400%
Day 10
77.3%
0.400%
Day 10
CD3+/CD8+
Day 10
77.3%
0.400%
Day 10
Day 10
Day 10
Day 10
Day 10
77.3%
77.3%
77.3%
77.3%
13.3%
0.408%
13.3%
13.3%
13.3%
13.3%
77.3%
6.60%
13.3%
2.37%
77.3%
6.60%
13.3%
2.37%
6.60%
6.60%
2.37%
2.37%
6.60%
6.60%
2.37%
2.37%
Day+/CD4
10 +
CD3
Day 10
Day 10
0.408%
0.408%
13.3%
0.408%
6.60%3.72%
2.37%
6.60%3.72%
2.37%
Day 10 +
CD3
Day+/CD4
10
3.72%
3.72%
0.529%
Day 14
Day 14
0.529%
0.529%
73.3%
0.529%
Day 14
Day 14 73.3%
Day 14
Day 14
0.529%
73.3%
73.3%
Day 14 73.3%
73.3%
Day 14
73.3%
11.5%
73.3%
11.5%
Day 14
Day 14
Day 14
Day 14
Day 14
Day 14
11.5%
11.5%
11.5%
11.5%
5.52%
11.5%
5.52%
11.5%
5.52%
5.52%
CD3
6.61%
0.019%
71.2%
Day 14 71.2%
CD3
CD3
Day 0
92.7% 0.447%
Day 0
1.31%
Day 14 71.2%
Day 14 71.2%
6.61%
0.019%
CD270.447%
9.59%
5.67%
5.67%
28.2%
28.2%
28.2%
0.408%
(D)
1.31%
1.31%
Day
0
Day
0
CD27
Day 10 83.6%
Day 10 83.6%
83.6%
0.400%
9.59%
9.59%
1.31%
15.8%
15.8%
Day 14 71.2%
1.31%
13.9%
9.59%
5.67%
Day
0
CD27
28.2%
28.2%
9.59%
82.0%
Day
0
CD27
Day
0
CD27
Day 14
Day 14 71.2%
83.6%
83.6%
1.31%
13.9%
Day
0
CD27
CD27
28.2%
Day 10
9.59%
82.0%
1.31%
1.31%
83.6%
Day 10
13.9%
13.9%
82.0%
82.0%
9.59%
9.59%
5.67%
Day 10
Day 10
15.8%
15.8%
Day 0
3.37%
Day
0
CD27
15.8%
15.8%
0.197%
0.019%
3.64%
CD3
Day 0
Day 14 71.2%
CD3
92.7%
92.7%
92.7%
3.37%
3.37%
CD27
CD57
5.67%
5.67%
5.67%
Day 0
CD57
CD57
Day
0
Day
0
CD57
CD57
Day 0
0
Day
CD57
Day 0
CD57
Day 0
Day 10
Day 10
13.3%
13.3%
6.60%
77.3%
13.3%
2.37%
6.60%
77.3%
13.3%
2.37%
6.60%
6.60%
2.37%
2.37%
Day 14
11.5%
11.5%
6.60%
6.60%
2.37%
2.37%
Day 14
11.5%
11.5%
3.72%
6.60%
2.37%
3.72%
6.60%
Day 14
14
Day
2.37%
Day
Day 14
14
Day+/CD4
10 +
CD3
Day 10
10
Day
Day 10
Day+/CD4
10 +
CD3
Day 10
3.72%
3.72%
3.72%
3.72%
Day 10
18.4%
18.4%
18.4%
18.4%
18.4%
18.4%
Day 14
73.3%
11.5%
73.3%
11.5%
5.52%
11.5%
5.52%
11.5%
5.52%
5.52%
Day 14
5.52%
5.52%
Day 14
3.72%
5.52%
3.72%
Day
10
Day+/CD8
10 +
CD3
Day 10
10
Day
Day+/CD8
10 +
CD3
Day 10
5.52%
Day 14
Day 10
18.4%
Day 14 73.3%
73.3%
77.3%
77.3%
Day 10
18.4%
CD57
Day 10
5.67%
Day 0
CD57
CD3
6.61%
6.61%
Day 0
Day 0
3.37%
3.37%
Day 0
CD3
92.7%
28.2%
CD3
CD3
6.61%
6.61%
92.7%
3.37%
3.64%
Day
0
CD27
CD3
CD3
(B)
Day 0
Day 0
92.7%
+
CD3
Day+/CD4
10 83.6%
CD3
CD28
CD28 CD28
CD28 CD28
CD28 CD28
CD28
6.61%
6.61%
15.8%
CD3
Day 0
3.37%
3.64%
3.37%
Day 14
CD3
Day 0
3.64%
3.64%
Day 14
2.27%
2.27%
2.27%
2.27%
2.27%
2.27%
2.27%
2.27%
Day
Day 14
14
Day 14
14
Day
Day 14
Day 14
3.82%
3.82%
3.82%
3.82%
3.82%
3.82%
3.82%
3.82%
CD57
CD57
CD57
CD57
CD57
CD57
CD57
CD3
CD3
CD3
CD3
CD3
CD3
CD3
+
+
+
+
Fig 3. Expression
of
well
Day
0 co-stimulatory molecules
Day 10 CD28 and CD27 as
Day
14 as CD57 in CD3 /CD4 T (helper) cells and CD3 /CD8 (cytotoxic) T cells in Xuri Cell
CD27
5.67%
5.67%
3.72%
5.52%
3.72%
5.52%
Expansion System
W5
at
day
0,
day
10
and
day
14
of
culture
(lymphocytes
were
gated
based
on
their
forwardand
side-scatter
profile). (A) Percentage
Day 0
Day 10
Day 14
5.67%
+
3.72%
5.52%
CD28+ and CD27+ within
CD3+/CD4+ T cells; (B) Percentage
CD28+ and CD27
within CD3+/CD8+ T cells. (C) Percentage CD57+ within CD3+/CD4+ T cells;
5.67% CD3+/CD8+ T cells. The expression patterns of the cell surface markers CD27 and CD28 give an indication of the differentiation
(D) Percentage CD57+ within
3.72%
5.52%
CD57
state of T cells. Naïve
or early differentiated T cells are CD27+/CD28+, effector T cells are CD27+/CD28- or CD27-/CD28+ and late effectors or ‘aged’ T cells are
Day 0
Day 1014 d of culture the majority
Day 14
CD27-/CD28- Expression
was high and after
of cells were in the early/intermediate stages of differentiation. CD57 expression
CD57
represents senescence
of
cells.
Flow
cytometric
analysis
showed
no
accumulation
of CD57+ T cells.
Day
0
Day
10
Day
14
CD57
Day 0
Day
0
CD57
Day 0
CD57
Day 0
18.4%
18.4%
18.4%
18.4%
18.4%
18.4%
18.4%
CD3
CD3
CD3
CD3
CD3
CD3
CD57
Day 0
Day
0
CD57
CD57
CD3
CD57
CD3
18.4%
CD57
CD57
2 29108247 AB
CD57
CD57
Day 10
2.27%
Day 14
Day 14
Day 10
Day 10
2.27%
Day 14
Day 14
Day+/CD8
10 +
CD3
2.27%
3.82%
3.82%
Day+/CD8
10 + 2.27%
CD3
Day 14
3.82%
3.82%
2.27%
2.27%
Day 14
3.82%
3.82%
Day 10
2.27%
2.27%
3.82%
3.82%
Xuri IL-15
Growth factors for research
Interleukin 15 (IL-15) is known to regulate activation and
proliferation of human blood lymphocytes. In cultivation
procedures it is commonly used to stimulate natural killer
(NK) cell growth.
IL-2
Xuri IL-15 includes information on biological activity in
reference to the international standard to allow consistent
performance over different batches and experiments.
• Minimized process development time
• Improved scale-out capacity
• Reproducible performance without revalidation of each lot
IL-2 enables cost-efficient expansion of T lymphocytes in
preclinical proof-of-principle experiments. Each lot of IL-2
includes information on biological activity to improve inter
batch reproducibility and save time on revalidation. The close
equivalence in production quality between IL-2 and Xuri IL-2
supports the transition of applications to clinical trial and
process development for cell therapy manufacturing with
minimal regulatory risk. The protocol provided with IL-2 was
specifically developed to support the expansion of T cells in
Xuri Cell Expansion Systems.
• Dedicated cell therapy ancillary product following
USP<1043> for ex vivo cultivation
IL-2 offers:
Xuri IL-15 supports compliance with USP<1043> (1). A
comprehensive product documentation support file is
available to facilitate risk assessment and validation for
use in cell therapy manufacturing.
• Accelerated setup time and reduced cost for transition
from research to the preclinical stage
Xuri IL-21
Interleukin 21 (IL-21) is recognized as being of significant
importance in the human immune system. Its role to regulate
NK cells and cytotoxic T cells renders it a key player in the
destruction of cancerous and virally infected cells.
• Reproducible performance without revalidation of each lot
• Quicker move to clinical stage through solid results
Further growth factors for research use are available
upon request.
Xuri IL-21 includes information on biological activity developed
in a functional assay against an internal standard. The clear
information on the activity allows batch-to-batch consistency
without time-consuming testing and revalidation.
• Minimized process development time
• Improved scale-out capacity
• Reproducible performance without revalidation of each lot
• Dedicated cell therapy ancillary product following
USP<1043> for ex vivo cultivation
Xuri IL-21 supports compliance with USP<1043> (1). A
comprehensive product documentation support file is
available to facilitate risk assessment and validation for
use in cell therapy manufacturing.
29108247 AB 3
Ordering information
Product
Pack size
Product code
Xuri IL-2 (USP<1043> for further
manufacturing only)
10 µg
29062789
Xuri IL-2 (USP<1043> for further
manufacturing only)
1 mg
29062790
Xuri IL-15 (USP<1043> for further
manufacturing only)
40 µg
29112116
Xuri IL-21 (USP<1043> for further
manufacturing only)
40 µg
29112119
IL-2 (For research use only)
10 µg
29062787
IL-2 (For research use only)
1 mg
29062788
www.gelifesciences.com/xuri
GE Healthcare UK Limited
Amersham Place
Little Chalfont
Buckinghamshire, HP7 9NA
UK
GE and GE monogram are trademarks of General Electric Company. Xuri and Cellbag are trademarks of General Electric Company or one of its subsidiaries.
© 2014–2015 General Electric Company―All rights reserved. First published Apr. 2014
Xuri 1L-2 and IL-2, Xuri IL-15 and Xuri IL-21 are neither IVDs or medicinal products and should not be used in diagnostic or therapeutic applications.
All goods and services are sold subject to the terms and conditions of sale of the company within GE Healthcare which supplies them. A copy of these
terms and conditions is available on request. Contact your local GE Healthcare representative for the most current information.
GE Healthcare Bio-Sciences AB, Björkgatan 30, 751 84 Uppsala, Sweden
GE Healthcare Europe, GmbH, Munzinger Strasse 5, D-79111 Freiburg, Germany
GE Healthcare Bio-Sciences Corp., 800 Centennial Avenue, P.O. Box 1327, Piscataway, NJ 08855-1327, USA
GE Healthcare Japan Corporation, Sanken Bldg., 3-25-1, Hyakunincho, Shinjuku-ku, Tokyo 169-0073, Japan
For local office contact information, visit www.gelifesciences.com/contact
29108247 AB 07/2015