Endothelial differentiation of bone marrow mesenchyme stem cells applicable to hypoxia and increased migration through Akt and NFκB signals.
Bone marrow mesenchymal stem cells (MSCs) and endothelial progenitor cells (EPCs) are used to repair hypoxic or ischemic tissue. However, the underlining mechanism of resistance in the hypoxic microenvironment and the efficacy of migration to the injured tissue are still unknown. The current study aims to understand the hypoxia resistance and migration ability of MSCs during differentiation toward endothelial lineages by biochemical and mechanical stimuli.



MSCs were harvested from the bone marrow of 6-8-week-old Sprague-Dawley rats. The endothelial growth medium (EGM) was added to MSCs for 3 days to initiate endothelial differentiation. Laminar shear stress was used as the fluid mechanical stimulation.



Application of EGM facilitated the early endothelial lineage cells (eELCs) to express EPC markers. When treating the hypoxic mimetic desferrioxamine, both MSCs and eELCs showed resistance to hypoxia as compared with the occurrence of apoptosis in rat fibroblasts. The eELCs under hypoxia increased the wound closure and C-X-C chemokine receptor type 4 (CXCR4) gene expression. Although the shear stress promoted eELC maturation and aligned cells parallel to the flow direction, their migration ability was not superior to that of eELCs either under normoxia or hypoxia. The eELCs showed higher protein expressions of CXCR4, phosphorylated Akt (pAkt), and endogenous NFκB and IκBα than MSCs under both normoxia and hypoxia conditions. The potential migratory signals were discovered by inhibiting either Akt or NFκB using specific inhibitors and revealed decreases of wound closure and transmigration ability in eELCs.



The Akt and NFκB pathways are important to regulate the early endothelial differentiation and its migratory ability under a hypoxic microenvironment.
Hyperbaric Oxygen Therapy Center, Chi-Mei Medical Center, Tainan, Taiwan.|Division of Plastic Surgery, Chi-Mei Medical Center, Tainan, Taiwan.|Department of Electrical Engineering, Southern Taiwan University of Science and Technology, Tainan, Taiwan.
Cheng
C
Liu
Division of Plastic Surgery, Chi-Mei Medical Center, Tainan, Taiwan.|Department of Cell Biology and Anatomy, College of Medicine, National Cheng Kung University, Tainan, Taiwan.
An-Ly
AL
Tsai
Department of Occupational Therapy, I-Shou University, Kaohsiung, Taiwan.|School of Medicine for International Students, I-Shou University, Kaohsiung, Taiwan.
Ping-Chia
PC
Li
Department of Cell Biology and Anatomy, College of Medicine, National Cheng Kung University, Tainan, Taiwan.|Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, Tainan, Taiwan.
Chia-Wei
CW
Huang
Department of Cell Biology and Anatomy, College of Medicine, National Cheng Kung University, Tainan, Taiwan. joshccwu@mail.ncku.edu.tw.|Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, Tainan, Taiwan. joshccwu@mail.ncku.edu.tw.|Medical Device Innovation Center, National Cheng Kung University, Tainan, Taiwan. joshccwu@mail.ncku.edu.tw.
Chia-Ching
0000-0001-7224-8425
CC
Wu
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D002454:Cell Differentiation / Q000187:drug effects; D015687:Cell Hypoxia / Q000235:genetics; D002465:Cell Movement / Q000187:drug effects; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology; D003676:Deferoxamine / Q000494:pharmacology; D066026:Endothelial Progenitor Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D005786:Gene Expression Regulation; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D000072000:NF-KappaB Inhibitor alpha / Q000235:genetics / Q000378:metabolism; D016328:NF-kappa B / Q000235:genetics* / Q000378:metabolism; D010100:Oxygen / Q000494:pharmacology*; D010766:Phosphorylation; D051057:Proto-Oncogene Proteins c-akt / Q000235:genetics* / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D019718:Receptors, CXCR4 / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D013314:Stress, Mechanical
D003470:Culture Media; C486264:Cxcr4 protein, rat; D016328:NF-kappa B; D019718:Receptors, CXCR4; D000072000:NF-KappaB Inhibitor alpha; D051057:Proto-Oncogene Proteins c-akt; D003676:Deferoxamine; D010100:Oxygen
Akt; Endothelial differentiation; Hypoxia; Migration; Nuclear factor-κB; Stem cell