Differentiation of Human Ligamentum Flavum Stem Cells Toward Nucleus Pulposus-Like Cells Induced by Coculture System and Hypoxia

Spine
Xiao-Bo HanChang-Qing Li

Abstract

Human ligamentum flavum (LF)-derived stem cells (LFSCs) and nucleus pulposus cells (NPCs) were cocultured under normoxia or hypoxia. To isolate and identify human LFSCs and determine whether they can differentiate into NPCs when cocultured with NPCs under hypoxia. Mesenchymal stem cell (MSC)-based therapies have been proposed as a biological treatment for intervertebral disc degeneration. MSCs derived from various tissues are leading candidates for cell-based therapies, but such cells have not been reported in LF. LF cells were isolated from patient samples and cultured using culture flasks coated with fibronectin, and their identity was confirmed using flow cytometry. The cells were induced to differentiate into osteoblasts, chondrocytes, and adipocytes, and their morphology, immunophenotype, cell proliferation capacity, cell cycle, and expression of stem cell-specific genes were compared with those of bone marrow-MSCs (BM-MSCs) derived from the same patients. NPCs and LFSCs were cocultured in 1-μm-pore-size insert transwell-culture systems under hypoxia (2% O2) or normoxia. CD24 expression was measured by flow cytometry and confocal microscopy assay. On day 14, reverse transcription-polymerase chain reaction was used for comp...Continue Reading

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Citations

Oct 1, 2016·Journal of Tissue Engineering and Regenerative Medicine·Naimisha BeeravoluG Rasul Chaudhry
Sep 9, 2017·Spine·Liam Harris, C Thomas Vangsness
Jan 2, 2019·Journal of Biological Engineering·Zhuochao LiuXingkai Zhang

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Methods Mentioned

BETA
FCS
PCR
confocal
flow cytometry
confocal microscopy

Software Mentioned

CellQuest

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