TY - JOUR
T1 - Phenotypic characteristics of human bone marrow-derived endothelial progenitor cells in vitro support cell effectiveness for repair of the blood-spinal cord barrier in ALS
AU - Garbuzova-Davis, Svitlana
AU - Ehrhart, Jared
AU - Mustafa, Hilmi
AU - Llauget, Alexander
AU - Boccio, Kayla J.
AU - Sanberg, Paul R.
AU - Appel, Stanley H.
AU - Borlongan, Cesario V.
N1 - Funding Information:
This study was supported by the NIH, NINDS (1R01NS090962) grant. SGD wrote the manuscript and analyzed data. JE performed ELISA, assisted with data analysis and designed the schematic diagram. HM assisted SGD with in vitro studies and performed immunocytochemistry, cell imaging, and ImageJ cell analyses. HM drew the schematic diagram. AL analyzed cell imaging for F-actin expressions and assisted HM in drawing the schematic. KJB assisted JE with performance of ELISA. SHA assisted SGD with study design and contributed to discussion of study results. CVB and PRS participated in topic discussions. All authors reviewed the manuscript. The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
Funding Information:
This study was supported by the NIH, NINDS ( 1R01NS090962 ) grant .
Publisher Copyright:
© 2019 Elsevier B.V.
PY - 2019/12/1
Y1 - 2019/12/1
N2 - Amyotrophic lateral sclerosis (ALS) was recently recognized as a neurovascular disease. Accumulating evidence demonstrated blood-spinal-cord barrier (BSCB) impairment mainly via endothelial cell (EC) degeneration in ALS patients and animal models. BSCB repair may be a therapeutic approach for ALS. We showed benefits of human bone marrow endothelial progenitor cell (hBMEPC) transplantation into symptomatic ALS mice on barrier restoration; however, cellular mechanisms remain unclear. The study aimed to characterize hBMEPCs in vitro under normogenic conditions. hBMEPCs were cultured at different time points. Enzyme-linked immunosorbent assay (ELISA) was used to detect concentrations of angiogenic factors (VEGF-A, angiogenin-1, and endoglin) and angiogenic inhibitor endostatin in conditioned media. Double immunocytochemical staining for CD105, ZO-1, and occludin with F-actin was performed. Results showed predominantly gradual significant post-culture increases of VEGF-A and angiogenin-1 levels. Cultured cells displayed distinct rounded or elongated cellular morphologies and positively immunoexpressed for CD105, indicating EC phenotype. Cytoskeletal F-actin filaments were re-arranged according to cell morphologies. Immunopositive expressions for ZO-1 were detected near inner cell membrane and for occludin on cell membrane surface of adjacent hBMEPCs. Together, secretion of angiogenic factors by cultured cells provides evidence for a potential mechanism underlying endogenous EC repair in ALS through hBMEPC transplantation, leading to restored barrier integrity. Also, ZO-1 and occludin immunoexpressions, confirming hBMEPC interactions in vitro, may reflect post-transplant cell actions in vivo.
AB - Amyotrophic lateral sclerosis (ALS) was recently recognized as a neurovascular disease. Accumulating evidence demonstrated blood-spinal-cord barrier (BSCB) impairment mainly via endothelial cell (EC) degeneration in ALS patients and animal models. BSCB repair may be a therapeutic approach for ALS. We showed benefits of human bone marrow endothelial progenitor cell (hBMEPC) transplantation into symptomatic ALS mice on barrier restoration; however, cellular mechanisms remain unclear. The study aimed to characterize hBMEPCs in vitro under normogenic conditions. hBMEPCs were cultured at different time points. Enzyme-linked immunosorbent assay (ELISA) was used to detect concentrations of angiogenic factors (VEGF-A, angiogenin-1, and endoglin) and angiogenic inhibitor endostatin in conditioned media. Double immunocytochemical staining for CD105, ZO-1, and occludin with F-actin was performed. Results showed predominantly gradual significant post-culture increases of VEGF-A and angiogenin-1 levels. Cultured cells displayed distinct rounded or elongated cellular morphologies and positively immunoexpressed for CD105, indicating EC phenotype. Cytoskeletal F-actin filaments were re-arranged according to cell morphologies. Immunopositive expressions for ZO-1 were detected near inner cell membrane and for occludin on cell membrane surface of adjacent hBMEPCs. Together, secretion of angiogenic factors by cultured cells provides evidence for a potential mechanism underlying endogenous EC repair in ALS through hBMEPC transplantation, leading to restored barrier integrity. Also, ZO-1 and occludin immunoexpressions, confirming hBMEPC interactions in vitro, may reflect post-transplant cell actions in vivo.
KW - Angiogenic factors
KW - F-actin
KW - Human bone marrow endothelial progenitor cells
KW - In vitro
KW - Tight junction proteins
UR - http://www.scopus.com/inward/record.url?scp=85072032542&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85072032542&partnerID=8YFLogxK
U2 - 10.1016/j.brainres.2019.146428
DO - 10.1016/j.brainres.2019.146428
M3 - Article
C2 - 31493389
AN - SCOPUS:85072032542
VL - 1724
JO - Brain Research
JF - Brain Research
SN - 0006-8993
M1 - 146428
ER -