Continuous harvest of stem cells via partial detachment from thermoresponsive nanobrush surfaces

Abstract

Stem cell culture is typically based on batch-type culture, which is laborious and expensive. Here, we propose a continuous harvest method for stem cells cultured on thermoresponsive nanobrush surfaces. In this method, stem cells are partially detached from the nanobrush surface by reducing the temperature of the culture medium below the critical solution temperature needed for thermoresponse. The detached stem cells are harvested by exchange into fresh culture medium. Following this, the remaining cells are continuously cultured by expansion in fresh culture medium at 37 °C. Thermoresponsive nanobrush surfaces were prepared by coating block copolymers containing polystyrene (for hydrophobic anchoring onto culture dishes) with three types of polymers: (a) polyacrylic acid with cell-binding oligopeptides, (b) thermoresponsive poly-N-isopropylacrylamide, and (c) hydrophilic poly(ethyleneglycol)methacrylate. The optimal coating durations and compositions for these copolymers to facilitate adequate attachment and detachment of human adipose-derived stem cells (hADSCs) and embryonic stem cells (hESCs) were determined. hADSCs and hESCs were continuously harvested for 5 and 3 cycles, respectively, via the partial detachment of cells from thermoresponsive nanobrush surfaces.

Full text

Data Article
Data of continuous harvest of stem cells
via partial detachment from thermoresponsive
nanobrush surfaces
Q2
Chin-Chen Yeh
a,1
, Saradaprasan Muduli
b,1
, I-Chia Peng
b,1
,
Yi-Tung Lu
b
, Qing-Dong Ling
c,d
, Abdullah A. Alarfaj
e
,
Murugan A. Munusamy
e
, S. Suresh Kumar
f
,
Kadarkarai Murugan
g
, Da-Chung Chen
h
, Hsin-chung Lee
i,j
,
Yung Chang
a,
n
, Akon Higuchi
b,e,
n
Q1
a
Department of Chemical Engineering, R&D Center for Membrane Technology, Chung Yuan Christian Uni-
versity, 200, Chung-Bei Rd., Chungli, Taoyuan 320, Taiwan
b
Department of Chemical and Materials Engineering, National Central University, No. 300, Jhongda Rd.,
Jhongli, Taoyuan 32001, Taiwan
c
Cathay Medical Research Institute, Cathay General Hospital, No. 32, Ln 160, Jian-Cheng Road, Hsi-Chi City,
Taipei 221, Taiwan
d
Institute of Systems Biology and Bioinformatics, National Central University, No. 300, Jhongda Rd., Jhongli,
Taoyuan 32001, Taiwan
e
Department of Botany and Microbiology, King Saud University, Riyadh 11451, Saudi Arabia
f
Department of Medical Microbiology and Parasitology, Universiti Putra Malaysia, Serdang 43400, Selangor,
Malaysia
g
Division of Entomology, Department of Zoology, School of Life Sciences, Bharathiar University, Coimbatore,
Tamil Nadu 641046, India
h
Department of Obstetrics and Gynecology, Taiwan Landseed Hospital, 77, Kuangtai Road, Pingjen City,
Taoyuan 32405, Taiwan
i
Department of Surgery, Cathay General Hospital, No.280, Sec. 4, Ren’ai Rd., Da’an Dist., Taipei 10693, Taiwan
j
Graduate Institute of Translational and Interdisciplinary Medicine, College of Health Science and Technology,
National Central University, No. 300, Jhongda Rd., Jhongli, Taoyuan 32001, Taiwan
article info
Article history:
Received 20 October 2015
Received in revised form
abstract
This data article contains two fi
Q3
gures and one table supporting the
research article entitled: “Continuous harvest of stem cells via partial
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Contents lists available at ScienceDirect
journal homepage: www.elsevier.com/locate/dib
Data in Brief
http://dx.doi.org/10.1016/j.dib.2015.12.056
2352-3409/&2016 Published by Elsevier Inc. This is an open access article under the CC BY license
(http://creativecommons.org/licenses/by/4.0/).
DOI of original article: http://dx.doi.org/10.1016/j.biomaterials.2015.10.039
n
Corresponding author.
n
Corresponding author at: Department of Chemical and Materials Engineering, National Central University, No. 300, Jhongda
Rd., Jhongli, Taoyuan 32001, Taiwan.
E-mail addresses: changyung0307@gmail.com (Y. Chang), higuchi@ncu.edu.tw (A. Higuchi).
1
These authors contributed equally to this work
Please cite this article as: C.-C. Yeh, et al., Data of continuous harvest of stem cells via partial
detachment from thermoresponsive nanobrush surfaces, Data in Brief (2016), http://dx.doi.org/
10.1016/j.dib.2015.12.056i
Data in Brief ∎(∎∎∎∎)∎∎∎–∎∎∎

12 December 2015
Accepted 28 December 2015
detachment from thermoresponsive nanobrush surface”[1].The
table shows coating conditions of three copolymers, poly(styrene-co-
acrylic acid) grafted with oligovitronectin, poly(styrene-co-N-iso-
propylacrylamide) and poly(styrene-co-polyethylene glycol metha-
crylate) to prepare thermoresponsive surface. XPS spectra show the
nitrogen peak of the polystyrene surface coated with poly(styrene-
co-acrylic acid) grafted with oligovitronectin. The surface coating
density analyzed from sorption of poly(styrene-co-acrylic acid) graf-
ted with oligovitronectin by UV–visspectroscopyisalsopresented.
&2016 Published by Elsevier Inc. This is an open access article under
the CC BY license (http://creativecommons.org/licenses/by/4.0/).
Specifications table
Subject area Chemistry
More specific sub-
ject area
Biomaterials
Type of data Table, figure
How data was
acquired
XPS, UV–vis spectroscopy
Data format Analyzed
Experimental
factors
Poly(styrene-co-acrylic acid) grafted with oligovitronectin was coated on
tissue culture polystyrene dishes
Experimental
features
See experimental details for each figure
Data source
location
Taiwan
Data accessibility Within this article
Value of the data

The data show coating conditions of three copolymers, poly(styrene-co-acrylic acid) grafted with
oligovitronectin, poly(styrene-co-N-isopropylacrylamide) and poly(styrene-co-polyethylene glycol
methacrylate) on polystyrene tissue culture plates for the preparation of thermoresponsive surface.

The data show which concentration of the coating polymer is necessary to cover the surface.

The surface coating density can be measured by spectroscopy on the surface coated with poly
(styrene-co-acrylic acid) grafted with oligovitronectin

The data show the evaluation of oligovitronectin measured by XPS spectra of the surface coated
with poly(styrene-co-acrylic acid) grafted with oligovitronectin

The existence of oligovitronectin on the surface coated with poly(styrene-co-acrylic acid) can be
verified by XPS measurement.
Data
Table 1 shows coating conditions of three copolymers, (a) poly(styrene-co-acrylic acid) grafted
with oligovitronectin (P[St-AA]-oligoVN), (b) poly(styrene-co-N-isopropylacrylamide) (P[St-PNI-
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Please cite this article as: C.-C. Yeh, et al., Data of continuous harvest of stem cells via partial
detachment from thermoresponsive nanobrush surfaces, Data in Brief (2016), http://dx.doi.org/
10.1016/j.dib.2015.12.056i
C.-C. Yeh et al. / Data in Brief ∎(∎∎∎∎)∎∎∎–∎∎∎2

PAAm]) and (c) poly(styrene-co-polyethylene glycol methacrylate) (P[St-PEGMA]) to prepare ther-
moresponsive surface.
Fig. 1 shows high-resolution X-ray photoelectron spectroscopy (XPS) spectra of the N1s peaks obtained
on the surface of 0% (a), 25% (b), 50% (c), 75% (d), and 100% (e) of surface coverage of P[St-AA]-oligoVN
where the surface coverage % of P[St-AA]-oligoVN is defined as % adsorption of [St-AA]-oligoVN on the
surface from the saturated adsorption amount (500 μg/cm
2
for 100%, 375 μg/cm
2
for 75%, 250 μg/cm
2
for
50%, and 125 μg/cm
2
for 25%). Nitrogen atoms originated from oligoVN on the surface [(b)-(e)], whereas no
nitrogen atoms were observed on the non-coated tissue culture polystyrene (TCPS) surface (a)
Fig. 2 shows dependence of surface coating density of P[St-AA]-oligoVN on the concentration of
coating solution. Coating density was measured by the decrease of optical density of coating solution
of P[St-AA]-oligoVN after immersion of TCPS plates into the coating solution.
1. Experimental design, materials and methods
We designed three types of coating copolymers: (a) a stem cell binding site, (b) a thermo-
responsive site, and (c) a hydrophilic site. Hydrophobic polystyrene (PSt) was selected as the
anchoring site of these three copolymers on the surface of TCPS. For this purpose, we synthesized
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Table 1
Coating conditions of thermoresponsive surface used in this study.
Coating conditions Coating concentration (mg/ml) Figure no. used in Ref. [1]
P[St-AA] P[St-PNIPAAm] P[St-PEG]
25% surface coverage of P[St-AA]-oligoVN 0.750 0 0 Fig. 4 A
25% surface coverage of P[St-AA]-oligoVN
with 1:0¼P[St-NIPAAm]: P[St-PEG]
0.750 3.000 0 Fig. 2A, 2D, 4A–D
25% surface coverage of P[St-AA]-oligoVN
with 9:1¼P[St-NIPAAm]: P[St-PEG]
0.750 2.700 0.300 Fig. 4B, 4 C, and 4D
25% surface coverage of P[St-AA]-oligoVN
with 4:1¼P[St-NIPAAm]: P[St-PEG]
0.750 2.400 0.600 Figs. 2A, C–E, 3, 4A–D
25% surface coverage of P[St-AA]-oligoVN
with 7:3¼P[St-NIPAAm]: P[St-PEG]
0.750 2.100 0.900 Fig. 2A and D
25% surface coverage of P[St-AA]-oligoVN
with 0:1¼P[St-NIPAAm]: P[St-PEG]
0.750 0 3.000 Fig. 2A, 4B–D
50% surface coverage of P[St-AA]-oligoVN
with 1:0¼P[St-NIPAAm]: P[St-PEG]
1.500 3.000 0 Fig. 4B–D
50% surface coverage of P[St-AA]-oligoVN
with 9:1¼P[St-NIPAAm]: P[St-PEG]
1.500 2.700 0.300 Fig. 4B–D
50% surface coverage of P[St-AA]-oligoVN
with 4:1¼P[St-NIPAAm]: P[St-PEG]
1.500 2.400 0.600 Fig. 4B–D
50% surface coverage of P[St-AA]-oligoVN
with 0:1¼P[St-NIPAAm]: P[St-PEG]
1.500 0 3.000 Fig. 4B–D
75% surface coverage of P[St-AA]-oligoVN
with 1:0¼P[St-NIPAAm]: P[St-PEG]
2.250 3.000 0 Fig. 4B–D
75% surface coverage of P[St-AA]-oligoVN
with 9:1¼P[St-NIPAAm]: P[St-PEG]
2.250 2.700 0.300 Fig. 4B–D
75% surface coverage of P[St-AA]-oligoVN
with 4:1¼P[St-NIPAAm]: P[St-PEG]
2.250 2.400 0.600 Fig. 4B–D
75% surface coverage of P[St-AA]-oligoVN
with 0:1¼P[St-NIPAAm]: P[St-PEG]
2.250 0 3.000 Fig. 4B–D
100% surface coverage of P[St-AA]-oligoVN
with 1:0¼P[St-NIPAAm]: P[St-PEG]
3.000 3.000 0 Fig. 4B–D
100% surface coverage of P[St-AA]-oligoVN
with 9:1¼P[St-NIPAAm]: P[St-PEG]
3.000 2.700 0.300 Fig. 4–D
100% surface coverage of P[St-AA]-oligoVN
with 4:1¼P[St-NIPAAm]: P[St-PEG]
3.000 2.400 0.600 Fig. 4–D, 5B–D, and 6
100% surface coverage of P[St-AA]-oligoVN
with 0:1¼P[St-NIPAAm]: P[St-PEG]
3.000 0 3.000 Fig. 4–D
Please cite this article as: C.-C. Yeh, et al., Data of continuous harvest of stem cells via partial
detachment from thermoresponsive nanobrush surfaces, Data in Brief (2016), http://dx.doi.org/
10.1016/j.dib.2015.12.056i
C.-C. Yeh et al. / Data in Brief ∎(∎∎∎∎)∎∎∎–∎∎∎ 3

three copolymers (a) P[St-AA]-oligoVN having the stem cell binding site of oligoVN (amino acid
sequence of KGGPQVTRGDVFTMP) [2], (b) P[St-NIPAAm] having thermoresponsive polyNIPAAm [3]
and (c) P[St-PEGMA] having hydrophilc PEGMA to prepare thermoresponsive surface.
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Fig. 2. Dependence of coating density of P[St-AA]-oligoVN on the concentration of coating solution. Coating density was mea-
sured by the decrease of optical density of coating solution of P[St-AA]-oligoVN after immersion of TCPS plates into the solution.
Fig. 1. High-resolution XPS spectra of the N1s peaks obtained on the surface of 0% (a), 25% (b), 50% (c), 75% (d), and 100% (e) of
surface coverage of P[St-AA]-oligoVN. Nitrogen atoms originated from oligoVN on the surface [(b)–(e)], whereas no nitrogen
atoms were observed on the non-coated TCPS surface (a).
Please cite this article as: C.-C. Yeh, et al., Data of continuous harvest of stem cells via partial
detachment from thermoresponsive nanobrush surfaces, Data in Brief (2016), http://dx.doi.org/
10.1016/j.dib.2015.12.056i
C.-C. Yeh et al. / Data in Brief ∎(∎∎∎∎)∎∎∎–∎∎∎4

1.1. Synthesis of copolymers
P[St-AA], P[St-NIPAAm], and P[St-PEGMA] were prepared by a reversible addition-fragmentation
chain transfer (RAFT) polymerization. The synthesis method of these copolymers was described in
Ref. [1] in detail.
1.2. Preparation process of thermoresponsive nanobrush surface
0–3 mg/mL of P[St-AA] in ethanol was added in TCPS dishes (4 cm
2
of surface area, 12 well dishes) for
coating of P[St-AA] on the surface for 2 h at 25 °C and subsequently removed from the dishes. TCPS dishes
coated with P[St-AA] were activated via immersion in an aqueous solution containing 10 mg/ml N-(3-dime-
thylaminopropyl)-N’-ethylcarbodiimide hydrochloride (EDC) and 10 mg/ml N-hydroxysuccinimide (NHS) for
1hat37°C after washing the dishes with phosphate buffered saline (PBS, pH 7.2) three times [1,4,5].Sub-
sequently, the dishes were washed with PBS and immersed in a PBS solution containing 1000 μg/mL of oli-
goVN for 24 h at 4 °C to prepare P[St-AA]-oligoVN dishes. The dishes were washed with PBS three times [1].
1.3. Characterization of dishes by XPS
The chemical composition of the dishes on the TCPS surface with P[St-AA]-oligoVN was analyzed
using X-ray photoelectron spectroscopy (XPS, K-Alpha spectrometer, Thermal Scientific, Inc., Amarillo,
TX, USA, equipped with a monochromatic Al-K X-ray source [1486.6 eV photons]). The energy of the
emitted electrons was measured using a hemispherical energy analyzer at pass energies ranging from
50 to 150 eV. Data were collected at a photoelectron takeoff angle of 45°with respect to the sample
surface. The binding energy (BE) scale was referenced by setting the peak maximum in the C1s
spectrum to 284.6 eV. The obtained high-resolution C1s spectra were fitted using Shirley background
subtraction and a series of Gaussian peaks [1,5].
Acknowledgements
This research was partially supported by the Ministry of Science and Technology under grant numbers
103-2120-M-008-001 and 104-2221-E-008-107-MY3. This work was also supported by the LandSeed
Hospital project (103LSH-NCU-1 and NCU-LSH-104-A-001) and the Cathay General Hospital Project
(103CGH-NCU-A3 and 104CGH-NCU-A3). The authors would like to extend their sincere appreciation to
the Deanship of Scientific Research at King Saud University for its funding of this research through the
Research Group project No RG-1435-065. A Grant-in-Aid for Scientific Research (15K06591) from the
Ministry of Education, Culture, Sports, Science, and Technology of Japan is also acknowledged.
Appendix A. Supplementary material
Supplementary data associated with this article can be found in the online version at http://dx.doi.
org/10.1016/j.dib.2015.12.056.
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Please cite this article as: C.-C. Yeh, et al., Data of continuous harvest of stem cells via partial
detachment from thermoresponsive nanobrush surfaces, Data in Brief (2016), http://dx.doi.org/
10.1016/j.dib.2015.12.056i
C.-C. Yeh et al. / Data in Brief ∎(∎∎∎∎)∎∎∎–∎∎∎6