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BD PuraMatrix Peptide Hydrogel
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Discovery Labware
Immunocytometry Systems
Pharmingen

BD
PuraMatrix
Peptide
Hydrogel
BD PuraMatrix Peptide
Hydrogel, a novel synthetic
matrix ideal for creating
optimized 3D cell culture
environments.
BD PuraMatrix Peptide Hydrogel is a
synthetic matrix that is used to create
defined three-dimensional (3D) microenvironments
for a variety of cell culture
experiments. To achieve optimal cell growth
and differentiation, it is necessary to
determine the appropriate mixture of this
material and bioactive molecules (e.g.,
growth factors, extracellular matrix
[ECM] proteins, and/or other molecules).
BD PuraMatrix Peptide Hydrogel consists
of standard amino acids (1% w/v) and
99% water. Under physiological conditions,
the peptide component self-assembles into
a 3D hydrogel that exhibits a nanometer
scale fibrous structure (Figure 1). The
hydrogel is readily formed in a culture dish,
plate, or cell culture insert.
The resulting hydrogel has been shown to
promote the differentiation of hepatocyte
progenitor cells 1 , rat pheochromocytoma
cells (PC12) 2 , and hippocampal neurons. 3
Studies have also demonstrated that
BD PuraMatrix Peptide Hydrogel supports
the attachment of a variety of primary (e.g.,
neuronal, fibroblast, keratinocyte) and
transformed (e.g., MG-63, SH-SY5Y,
HEK293, NIH3T3) cell types. 4 Other
potential applications include stem cell
proliferation, tumor cell migration and
invasion, angiogenesis assays, and in vivo
analyses of tissue regeneration.
BD PuraMatrix Peptide Hydrogel is
biocompatible, resorbable, and devoid of
animal-derived material and pathogens. For
in vivo studies in animals, the soluble
material can be injected and will subsequently
form a 3D hydrogel upon contact
with the physiological environment.
Application Focus
Differentiation of Hepatocyte Progenitor Cells 1
Rat hepatocyte progenitor cells (Lig-8 5 ) were encapsulated in BD PuraMatrix Peptide
Hydrogel and cultured overnight in defined medium at 37ºC. Samples were then used for
bromodioxyuridine (BrdU) uptake and in situ immunofluorescence analyses. As shown in
Figure 2, Lig-8 cells form spheroid colonies when cultured within the 3D hydrogel and
express the hepatocyte markers CCAAT/enhancer binding protein α (C/EBPα) and cytochrome
P450 1A1/1A2 (CYP1A1/1A2) in a manner that is independent of cellular mitotic
activity. Therefore, while some cells are proliferating, the entire colony exhibits
differentiation potential.
Figure 1. Electron micrograph of BD PuraMatrix
Peptide Hydrogel [bar, 100 nm].
Figure 2. Lig-8 cells cultured in BD PuraMatrix
Peptide Hydrogel. All cells in spheroid colonies,
arrested or not, undergo differentiation.
Spheroids were isolated, transferred to
adherent cultures, and incubated with BrdU
for 24 hours.
(A) spheroid colony (phase contrast)
(B) same optical layer as A immunostained for
C/EBPα (red)
References
1. Semino, C.E., et al., Functional differentiation
of hepatocyte-like spheroid structures from
putative liver progenitor cells in threedimensional
peptide scaffolds. Differentiation
71:262 (2003).
2. Holmes, T.C., et al., Extensive neurite outgrowth
and active synapse formation on selfassembling
peptide scaffolds. PNAS USA
97:6728 (2000).
3. Semino, C.E., et al., Entrapment of migrating
hippocampal neural cells in 3D peptide nanofiber
scaffold. Tissue Engineering 10:643 (2004).
4. Zhang, S., et al., Self-complementary oligopeptide
matrices support mammalian cell
attachment. Biomaterials 16:1385 (1995).
5. Lee, H-S., et al., Clonal expansion of adult rat
hepatic stem cell lines by suppression of
asymmetric cell kinetics (SACK). Biotechnology
and Bioengineering 83:760 (2003).
(C) same optical layer as A immunostained for
BrdU (green)
(D) spheroid colony (phase contrast)
(E) same optical layer as D immunostained for
CYP1A1/1A2 (red)
(F) same optical layer as D immunostained for
BrdU
Data provided by 3D Matrix, Inc. and originally
described in Semino, C.E., et al., Differentiation
71:262 (2003).

Analysis of Neurite Outgrowth using PC12 cells 2
PC12 cells were cultured on BD PuraMatrix Peptide Hydrogel according to a surface plating protocol. To examine the differentiated
morphology of these cells, samples were incubated in the presence of nerve growth factor (NGF) and subsequently analyzed by scanning
laser confocal microscopy. PC12 cells cultured under these conditions were found to differentiate and exhibit pronounced neurite outgrowth
(Figure 3). Moreover, human SY5Y neuroblastoma cells and a variety of primary neurons were shown to exhibit comparable
activity on this material (Table 1).
Table 1. Neurite outgrowth from neuronal cells on BD PuraMatrix Peptide Hydrogel.
Cell Type Neurite Length, µm Cell Source†
NGF-treated Rat PC12 400-500 Cultured cell line
NGF-preprimed PC12 400-500 Cultured cell line
Human SY5Y neuroblastoma 400-500 Cultured cell line
Mouse cerebellar granule neurons 200-300 Primary cells‡
Mouse hippocampal neurons 100-200 Primary cells‡
Rat hippocampal neurons 200-300 Primary cells§
Figure 3. PC12 cell neurite outgrowth and
differentiation on BD PuraMatrix Peptide
Hydrogel. The image is a merged stack of
multiple confocal optical sections. PC12 cells
cultured in the presence of NGF attached to the
hydrogel and projected extensive neurites. The
black spots are holes in the hydrogel. The
micrograph is representative of at least four
independent experiments.
† Cells were seeded onto BD PuraMatrix Peptide Hydrogel. The cell-bearing hydrogels were transferred
to dishes with fresh medium. Maximum neurite length was estimated visually with scale bars 3 to 7
days after cell attachment for primary cells and 10 to 14 days for the cultured cell lines.
‡ Seven-day-old mouse
§ One-day-old rat
Data provided by 3D Matrix, Inc. and originally
described in Holmes, T.C., et al., PNAS USA 97:
6728 (2000).
Differentiation of Neurons Derived from Hippocampal Slices 3
Hippocampal tissue slices (~200 µm
thickness) were isolated from 7.5-day-old
postnatal rats and placed on top of a
microporous membrane insert for up to
two weeks. The inserts were either
uncoated (control samples) or coated
with BD PuraMatrix Peptide Hydrogel
(~500 µm thick). When cultured on the
hydrogel, the hippocampal slice exhibited
tissue growth and cell migration into the
material (Figure 4). While neuronal
migration was not observed in the control
samples (Figure 4g), neuronal and glial
progenitor cells were found to migrate
from the tissue slice into the hydrogel layer
(Figure 4f and 4h). These results suggest
that BD PuraMatrix Peptide Hydrogel
may be useful for promoting neural cell
differentiation and tissue growth in vivo.
Figure 4. Hippocampal organotypic slice
cultures on BD PuraMatrix Peptide Hydrogel
develop extended tissue growth. Hippocampal
slices were cultured on control
membrane or BD PuraMatrix Peptide
Hydrogel layers (~500 µm thick). Time lapse
was carried out to follow tissue growth
from the perimeter of the dentate gyrus
region. (A) Time 0 (0 hour) of control slice
culture; (B) Time 0 (0 hour) hydrogel
(RAD16-I) slice culture; (C) 72 hours,
control slice; (D) 72 hours, hydrogel slice;
The red line indicates the original border
of the tissue slice and the yellow line in d
shows the extended tissue growth. The
yellow arrow in d indicates the direction
of tissue growth and extension. Black bars
represent 100 µm. (E) 72 hours, control
slice immunostained for GFAP (glial cell
marker, green); (F) 72 hours, hydrogel
slice immunostained for GFAP (green);
(G) same optical layer as e immunostained
for NeuN (neuron marker, red); (H) same
optical layer as f immunostained for NeuN
(red). Red lines in e and f and yellow in g
and h indicate original perimeter of the
tissue slice. The white line in e and g indicates
the extended tissue scaffold in control
cultures. The white line in f and h is
used to compare the over extension
obtained on hydrogel cultures. Yellow
arrows in h indicate NeuN+ neurons (red)
migrating into Area II in hydrogel slice cultures.
White bar in f represents 100 µm.
Data provided by 3D Matrix, Inc. and
originally described in Semino, C.E., et al.,
Tissue Engineering 10:643 (2004).

BD PuraMatrix Peptide Hydrogel
Features and Benefits
Purified Synthetic Peptide
Composition (1% w/v)
3D Hydrogel Structure
Easy Handling
Transparent Hydrogel
Established Protocols
Highly defined material that promotes cell attachment
Assembles into fibrous structure with average pore size of 50-200 nm
Easily mixed with cells and/or bioactive molecules (e.g., growth factors) prior to gelation;
injectable for in vivo studies in animals
Samples are readily visualized using standard staining methodologies and microscopy
3D cell encapsulation cultures; Surface plating of adherent cells on microporous membrane
inserts and microplates; cell recovery for sub-culturing or biochemical analyses; in vivo
injection
Characteristics
• Peptide sequence promotes cell attachment, but does not activate RGD-dependent integrin signaling
• In the presence of salt-containing solution, the peptide component of BD PuraMatrix Peptide Hydrogel
self-assembles and forms a transparent 3D hydrogel
• Exhibits nanometer scale fibrous structure
• Biocompatible; devoid of animal-derived material and pathogens
Technical Specifications
• 1% solution (w/v) of purified synthetic peptide
• Packaged material exhibits pH = 3.0
• Quality Control:
– Tested and found negative for bacteria, fungi, and Mycoplasma
– Cell viability > 80% based on cytotoxicity analysis of NIH3T3 fibroblasts
– Identity confirmed using Mass Spectrometry
– Demonstration of fiber formation using a self-assembly assay
Ordering Information
Description Qty Cat. No.
BD PuraMatrix Peptide Hydrogel 5 ml 354250
To place an order in the U.S., contact Customer Service at:
tel: 800.343.2035 or 978.901.7300; fax: 800.743.6200 or 978.901.7493
For technical assistance, contact Technical Service at:
tel: 800.343.2035 or 978.901.7300; fax: 800.743.6200 or 978.901.7493
To place an order outside the U.S., contact your local distributor or nearest
BD Biosciences office.
For information about GMP-grade material, contact 3D Matrix, Inc. at:
e-mail: sales@puramatrix.com; internet: www.puramatrix.com
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For Research Use Only. Not for use in diagnostic or therapeutic procedures. Not for resale.
PuraMatrix is a registered trademark of 3D Matrix, Inc.
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