TSP-1 secreted by one marrow stroma cells contributes to retinal ganglion cell neurite outgrowth and survival

Keming Yu, Jian Ge, James Bradley Summers, Fan Li, Xuan Liu, Ping Ma, Joseph Michael Kaminski, Jing Zhuang

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

Background: Bone marrow stromal cells (BMSCs) are pluripotent and thereby a potential candidate for cell replacement therapy for central nervous system degenerative disorders and traumatic injury. However, the mechanism of their differentiation and effect on neural tissues has not been fully elucidated. This study evaluates the effect of BMSCs on neural cell growth and survival in a retinal ganglion cell (RGCs) model by assessing the effect of changes in the expression of a BMSC-secreted protein, thrombospondin-1 (TSP-1), as a putative mechanistic agent acting on RGCs. Methods and Findings: The effect of co-culturing BMSCs and RGCs in vitro was evaluated by measuring the following parameters: neurite outgrowth, RGC survival, BMSC neural-like differentiation, and the effect of TSP-1 on both cell lines under basal secretion conditions and when TSP-1 expression was inhibited. Our data show that BMSCs improved RGC survival and neurite outgrowth. Synaptophysin, MAP-2, and TGF-β expression are up-regulated in RGCs co-cultured with BMSCs. Interestingly, the BMSCs progressively displayed neural-like morphology over the seven-day study period. Restriction display polymerase chain reaction (RD-PCR) was performed to screen for differentially expressed genes in BMSCs cultured alone or cocultured with RGCs. TSP-1, a multifactorial extracellular matrix protein, is critically important in the formation of neural connections during development, so its function in our co-culture model was investigated by small interfering RNA (siRNA) transfection. When TSP-1 expression was decreased with siRNA silencing, BMSCs had no impact on RGC survival, but reduced neurite outgrowth and decreased expression of synaptophysin, MAP-2 and TGF-β in RGCs. Furthermore, the number of BMSCs with neural-like characteristics was significantly decreased by more than two-fold using siRNA silencing. Conclusions: Our data suggest that the TSP-t signaling pathway might have an important role in neural-like differentiation in BMSCs and neurite outgrowth in RGCs. This study provides new insights into the potential reparative mechanisms of neural cell repair.

Original languageEnglish (US)
Article numbere2470
JournalPloS one
Volume3
Issue number6
DOIs
StatePublished - Jun 25 2008

Fingerprint

Thrombospondin 1
neurites
Retinal Ganglion Cells
stromal cells
bone marrow cells
Mesenchymal Stromal Cells
Bone
Bone Marrow
cells
Cells
cell viability
Cell Survival
small interfering RNA
Small Interfering RNA
Synaptophysin
RNA Interference
RNA interference
Neuronal Outgrowth
neurons
Extracellular Matrix Proteins

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

TSP-1 secreted by one marrow stroma cells contributes to retinal ganglion cell neurite outgrowth and survival. / Yu, Keming; Ge, Jian; Summers, James Bradley; Li, Fan; Liu, Xuan; Ma, Ping; Kaminski, Joseph Michael; Zhuang, Jing.

In: PloS one, Vol. 3, No. 6, e2470, 25.06.2008.

Research output: Contribution to journalArticle

Yu, K, Ge, J, Summers, JB, Li, F, Liu, X, Ma, P, Kaminski, JM & Zhuang, J 2008, 'TSP-1 secreted by one marrow stroma cells contributes to retinal ganglion cell neurite outgrowth and survival', PloS one, vol. 3, no. 6, e2470. https://doi.org/10.1371/journal.pone.0002470
Yu, Keming ; Ge, Jian ; Summers, James Bradley ; Li, Fan ; Liu, Xuan ; Ma, Ping ; Kaminski, Joseph Michael ; Zhuang, Jing. / TSP-1 secreted by one marrow stroma cells contributes to retinal ganglion cell neurite outgrowth and survival. In: PloS one. 2008 ; Vol. 3, No. 6.
@article{40dd20d7d55b43f48c2216bbcb38f605,
title = "TSP-1 secreted by one marrow stroma cells contributes to retinal ganglion cell neurite outgrowth and survival",
abstract = "Background: Bone marrow stromal cells (BMSCs) are pluripotent and thereby a potential candidate for cell replacement therapy for central nervous system degenerative disorders and traumatic injury. However, the mechanism of their differentiation and effect on neural tissues has not been fully elucidated. This study evaluates the effect of BMSCs on neural cell growth and survival in a retinal ganglion cell (RGCs) model by assessing the effect of changes in the expression of a BMSC-secreted protein, thrombospondin-1 (TSP-1), as a putative mechanistic agent acting on RGCs. Methods and Findings: The effect of co-culturing BMSCs and RGCs in vitro was evaluated by measuring the following parameters: neurite outgrowth, RGC survival, BMSC neural-like differentiation, and the effect of TSP-1 on both cell lines under basal secretion conditions and when TSP-1 expression was inhibited. Our data show that BMSCs improved RGC survival and neurite outgrowth. Synaptophysin, MAP-2, and TGF-β expression are up-regulated in RGCs co-cultured with BMSCs. Interestingly, the BMSCs progressively displayed neural-like morphology over the seven-day study period. Restriction display polymerase chain reaction (RD-PCR) was performed to screen for differentially expressed genes in BMSCs cultured alone or cocultured with RGCs. TSP-1, a multifactorial extracellular matrix protein, is critically important in the formation of neural connections during development, so its function in our co-culture model was investigated by small interfering RNA (siRNA) transfection. When TSP-1 expression was decreased with siRNA silencing, BMSCs had no impact on RGC survival, but reduced neurite outgrowth and decreased expression of synaptophysin, MAP-2 and TGF-β in RGCs. Furthermore, the number of BMSCs with neural-like characteristics was significantly decreased by more than two-fold using siRNA silencing. Conclusions: Our data suggest that the TSP-t signaling pathway might have an important role in neural-like differentiation in BMSCs and neurite outgrowth in RGCs. This study provides new insights into the potential reparative mechanisms of neural cell repair.",
author = "Keming Yu and Jian Ge and Summers, {James Bradley} and Fan Li and Xuan Liu and Ping Ma and Kaminski, {Joseph Michael} and Jing Zhuang",
year = "2008",
month = "6",
day = "25",
doi = "10.1371/journal.pone.0002470",
language = "English (US)",
volume = "3",
journal = "PLoS One",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "6",

}

TY - JOUR

T1 - TSP-1 secreted by one marrow stroma cells contributes to retinal ganglion cell neurite outgrowth and survival

AU - Yu, Keming

AU - Ge, Jian

AU - Summers, James Bradley

AU - Li, Fan

AU - Liu, Xuan

AU - Ma, Ping

AU - Kaminski, Joseph Michael

AU - Zhuang, Jing

PY - 2008/6/25

Y1 - 2008/6/25

N2 - Background: Bone marrow stromal cells (BMSCs) are pluripotent and thereby a potential candidate for cell replacement therapy for central nervous system degenerative disorders and traumatic injury. However, the mechanism of their differentiation and effect on neural tissues has not been fully elucidated. This study evaluates the effect of BMSCs on neural cell growth and survival in a retinal ganglion cell (RGCs) model by assessing the effect of changes in the expression of a BMSC-secreted protein, thrombospondin-1 (TSP-1), as a putative mechanistic agent acting on RGCs. Methods and Findings: The effect of co-culturing BMSCs and RGCs in vitro was evaluated by measuring the following parameters: neurite outgrowth, RGC survival, BMSC neural-like differentiation, and the effect of TSP-1 on both cell lines under basal secretion conditions and when TSP-1 expression was inhibited. Our data show that BMSCs improved RGC survival and neurite outgrowth. Synaptophysin, MAP-2, and TGF-β expression are up-regulated in RGCs co-cultured with BMSCs. Interestingly, the BMSCs progressively displayed neural-like morphology over the seven-day study period. Restriction display polymerase chain reaction (RD-PCR) was performed to screen for differentially expressed genes in BMSCs cultured alone or cocultured with RGCs. TSP-1, a multifactorial extracellular matrix protein, is critically important in the formation of neural connections during development, so its function in our co-culture model was investigated by small interfering RNA (siRNA) transfection. When TSP-1 expression was decreased with siRNA silencing, BMSCs had no impact on RGC survival, but reduced neurite outgrowth and decreased expression of synaptophysin, MAP-2 and TGF-β in RGCs. Furthermore, the number of BMSCs with neural-like characteristics was significantly decreased by more than two-fold using siRNA silencing. Conclusions: Our data suggest that the TSP-t signaling pathway might have an important role in neural-like differentiation in BMSCs and neurite outgrowth in RGCs. This study provides new insights into the potential reparative mechanisms of neural cell repair.

AB - Background: Bone marrow stromal cells (BMSCs) are pluripotent and thereby a potential candidate for cell replacement therapy for central nervous system degenerative disorders and traumatic injury. However, the mechanism of their differentiation and effect on neural tissues has not been fully elucidated. This study evaluates the effect of BMSCs on neural cell growth and survival in a retinal ganglion cell (RGCs) model by assessing the effect of changes in the expression of a BMSC-secreted protein, thrombospondin-1 (TSP-1), as a putative mechanistic agent acting on RGCs. Methods and Findings: The effect of co-culturing BMSCs and RGCs in vitro was evaluated by measuring the following parameters: neurite outgrowth, RGC survival, BMSC neural-like differentiation, and the effect of TSP-1 on both cell lines under basal secretion conditions and when TSP-1 expression was inhibited. Our data show that BMSCs improved RGC survival and neurite outgrowth. Synaptophysin, MAP-2, and TGF-β expression are up-regulated in RGCs co-cultured with BMSCs. Interestingly, the BMSCs progressively displayed neural-like morphology over the seven-day study period. Restriction display polymerase chain reaction (RD-PCR) was performed to screen for differentially expressed genes in BMSCs cultured alone or cocultured with RGCs. TSP-1, a multifactorial extracellular matrix protein, is critically important in the formation of neural connections during development, so its function in our co-culture model was investigated by small interfering RNA (siRNA) transfection. When TSP-1 expression was decreased with siRNA silencing, BMSCs had no impact on RGC survival, but reduced neurite outgrowth and decreased expression of synaptophysin, MAP-2 and TGF-β in RGCs. Furthermore, the number of BMSCs with neural-like characteristics was significantly decreased by more than two-fold using siRNA silencing. Conclusions: Our data suggest that the TSP-t signaling pathway might have an important role in neural-like differentiation in BMSCs and neurite outgrowth in RGCs. This study provides new insights into the potential reparative mechanisms of neural cell repair.

UR - http://www.scopus.com/inward/record.url?scp=49649084238&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=49649084238&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0002470

DO - 10.1371/journal.pone.0002470

M3 - Article

C2 - 18575624

AN - SCOPUS:49649084238

VL - 3

JO - PLoS One

JF - PLoS One

SN - 1932-6203

IS - 6

M1 - e2470

ER -