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IUBMB Life. 2019 May 21. doi: 10.1002/iub.2074. [Epub ahead of print]

A comparative study of HAMSCs/HBMSCs transwell and mixed coculture systems.

Bian Y1, Du Y1,2, Wang R1,3, Chen N1,2, Du X4, Wang Y1,2, Yuan H1,2.

Author information

1
Jiangsu Key Laboratory of Oral Diseases, Nanjing Medical University, Nanjing, China.
2
Department of Oral and Maxillofacial Surgery, Affiliated Hospital of Stomatology, Nanjing Medical University, Nanjing, China.
3
Department of Dental Implant, Affiliated Hospital of Stomatology, Nanjing Medical University, Nanjing, China.
4
State Key Laboratory of Bioelectronics, Southeast University, Nanjing, China.

Abstract

Our previous studies indicated that a coculture system containing human amnion-derived mesenchymal stem cells (HAMSCs) and human bone marrow mesenchymal stem cells (HBMSCs) has the potential of application for bone regeneration. However, there is currently no enough comparative investigation between HAMSCs/HBMSCs transwell and mixed coculture systems. This study aimed to assess the phenotype and mechanisms regulated by indirect and direct coculture systems, respectively. Two in vitro models were employed with HAMSCs and HBMSCs at a ratio of 3:1, and then were analyzed by a series of processes, including flow cytometry, alkaline phosphatase (ALP) substrate assays, Alizarin red S staining, quantitative reverse transcription polymerase chain reaction (RT-qPCR), and Western blot analysis. We found that cell proliferation, ALP activity, mineralized matrix formation, and osteoblast-related mRNA expression were accelerated in transwell coculture system compared with mixed coculture system. Conditioned medium from transwell coculture system achieved an elevated level of vascular endothelial growth factor and induced more vascular structures in human umbilical vein endothelial cells than those of mixed coculture system. Moreover, we observed that transwell coculture system, promoted osteogenesis and angiogenesis by maintaining stemness through extracellular regulated protein kinases 1/2 (ERK1/2) mitogen-activated protein kinase (MAPK) signaling pathway. U0126, a selective inhibitor of ERK1/2 MAPK signaling, significantly suppressed maintaining of the stemness-based effects on transwell coculture system. Taken together, our results compared the merits of two different models and clarified the role of HAMSCs/HBMSCs transwell coculture system in the development of bone tissue engineering.

KEYWORDS:

coculture system; human amnion-derived mesenchymal stem cells; human bone marrow mesenchymal stem cells; mitogen-activated protein kinase signaling pathway; stemness

PMID:
31112365
DOI:
10.1002/iub.2074

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