Accepted Manuscript Effects of hypoxia on differentiation from human placenta derived mesenchymal stem cells to nucleus pulposus-like cells Li Ni, MD Xiaochen Liu, BS Kyle R. Sochacki, BS Miranda Ebraheim, BS Matthew Fahrenkopf, BS Qin Shi, MD Jiayong Liu, MD Huilin Yang, MD, PhD PII:
S1529-9430(14)00302-7
DOI:
10.1016/j.spinee.2014.03.028
Reference:
SPINEE 55823
To appear in:
The Spine Journal
Received Date: 2 July 2013 Revised Date:
8 March 2014
Accepted Date: 16 March 2014
Please cite this article as: Ni L, Liu X, Sochacki KR, Ebraheim M, Fahrenkopf M, Shi Q, Liu J, Yang H, Effects of hypoxia on differentiation from human placenta derived mesenchymal stem cells to nucleus pulposus-like cells, The Spine Journal (2014), doi: 10.1016/j.spinee.2014.03.028. This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
ACCEPTED MANUSCRIPT
Effects of hypoxia on differentiation from human placenta derived
RI PT
mesenchymal stem cells to nucleus pulposus-like cells
Li Ni1 MD; Xiaochen Liu2 BS; Kyle R. Sochacki2 BS; Miranda Ebraheim2 BS; Matthew Fahrenkopf 2 BS; Qin Shi1 MD; Jiayong Liu 1, 2 MD; Huilin Yang1 MD, PhD The First Affiliated Hospital of Suzhou University, Suzhou, 215006, China
2
University of Toledo Medical Center, Toledo, OH 43614, USA.
M AN U
SC
1
TE D
Correspondence to: Huilin Yang MD, PhD Department of Orthopaedic Surgery The First Affiliated Hospital of Soochow University 188 Shizi Street, Suzhou, Jiangsu 215006, China Tel: +86-512-67780101 Fax: +86-512-67780999 E-mail address: [email protected]
AC C
EP
Co-correspondence to: Jiayong Liu MD Department of Orthopaedic Surgery University of Toledo Medical Center 3065 Arlington Avenue Toledo, Ohio, USA 43614 Phone: 419-383-5361 Fax: 419-383-3526 E-mail: [email protected]
This paper has been orally presented at North American Spine Society 2012 (27th) Annual Meeting, October 24-27, 2012, Dallas, TX.
ACCEPTED MANUSCRIPT
Abstract
1
Background: Low back pain is a frequently occurring disease caused by intervertebral disc
3
degeneration. Mesenchymal stem cells (MSCs) are a possible treatment modality. Studies have
4
shown MSCs can be transformed into nucleus pulposus-like cells under normoxic conditions.
5
However, this is not a true representation of the hypoxic environment nucleus pulposus cells
6
experience during in vivo growth and differentiation.
7
Purpose: To determine the effects of a hypoxic environment on the differentiation of human
8
placenta derived mesenchymal stem cells (PMSCs) to nucleus pulposus-like cells.
9
Study Design: An experimental study.
M AN U
SC
RI PT
2
Methods: PMSCs were cultured and the mesenchymal lineage was confirmed by flow
11
cytometry. Two groups of PMSCs were then cultured under different oxygen concentrations
12
creating a hypoxic group and normoxic group. The proliferation of cells in each group was
13
compared by CCK-8 on day 1, 3, 5, and 7. RT-PCR on days 3 and 7 compared the expressions of
14
Sox-9, type II collagen, aggrecan, and HIF-1α between the two groups. Immunofluorescence was
15
used to compare the expression of type II collagen between the two groups of after 14 days.
16
Results: PMSCs were successfully isolated and cultured. Mesenchymal markers were positive.
17
On day 3 and 5, the hypoxic group had a significantly higher proliferation rate than the normoxic
18
group (p
S1529-9430(14)00302-7
DOI:
10.1016/j.spinee.2014.03.028
Reference:
SPINEE 55823
To appear in:
The Spine Journal
Received Date: 2 July 2013 Revised Date:
8 March 2014
Accepted Date: 16 March 2014
Please cite this article as: Ni L, Liu X, Sochacki KR, Ebraheim M, Fahrenkopf M, Shi Q, Liu J, Yang H, Effects of hypoxia on differentiation from human placenta derived mesenchymal stem cells to nucleus pulposus-like cells, The Spine Journal (2014), doi: 10.1016/j.spinee.2014.03.028. This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
ACCEPTED MANUSCRIPT
Effects of hypoxia on differentiation from human placenta derived
RI PT
mesenchymal stem cells to nucleus pulposus-like cells
Li Ni1 MD; Xiaochen Liu2 BS; Kyle R. Sochacki2 BS; Miranda Ebraheim2 BS; Matthew Fahrenkopf 2 BS; Qin Shi1 MD; Jiayong Liu 1, 2 MD; Huilin Yang1 MD, PhD The First Affiliated Hospital of Suzhou University, Suzhou, 215006, China
2
University of Toledo Medical Center, Toledo, OH 43614, USA.
M AN U
SC
1
TE D
Correspondence to: Huilin Yang MD, PhD Department of Orthopaedic Surgery The First Affiliated Hospital of Soochow University 188 Shizi Street, Suzhou, Jiangsu 215006, China Tel: +86-512-67780101 Fax: +86-512-67780999 E-mail address: [email protected]
AC C
EP
Co-correspondence to: Jiayong Liu MD Department of Orthopaedic Surgery University of Toledo Medical Center 3065 Arlington Avenue Toledo, Ohio, USA 43614 Phone: 419-383-5361 Fax: 419-383-3526 E-mail: [email protected]
This paper has been orally presented at North American Spine Society 2012 (27th) Annual Meeting, October 24-27, 2012, Dallas, TX.
ACCEPTED MANUSCRIPT
Abstract
1
Background: Low back pain is a frequently occurring disease caused by intervertebral disc
3
degeneration. Mesenchymal stem cells (MSCs) are a possible treatment modality. Studies have
4
shown MSCs can be transformed into nucleus pulposus-like cells under normoxic conditions.
5
However, this is not a true representation of the hypoxic environment nucleus pulposus cells
6
experience during in vivo growth and differentiation.
7
Purpose: To determine the effects of a hypoxic environment on the differentiation of human
8
placenta derived mesenchymal stem cells (PMSCs) to nucleus pulposus-like cells.
9
Study Design: An experimental study.
M AN U
SC
RI PT
2
Methods: PMSCs were cultured and the mesenchymal lineage was confirmed by flow
11
cytometry. Two groups of PMSCs were then cultured under different oxygen concentrations
12
creating a hypoxic group and normoxic group. The proliferation of cells in each group was
13
compared by CCK-8 on day 1, 3, 5, and 7. RT-PCR on days 3 and 7 compared the expressions of
14
Sox-9, type II collagen, aggrecan, and HIF-1α between the two groups. Immunofluorescence was
15
used to compare the expression of type II collagen between the two groups of after 14 days.
16
Results: PMSCs were successfully isolated and cultured. Mesenchymal markers were positive.
17
On day 3 and 5, the hypoxic group had a significantly higher proliferation rate than the normoxic
18
group (p
