Effects of spheroid culture in a rotating bioreactor on secretion of inflammatory factors by human placenta-derived mesenchymal stem cells

doi:10.12307/2024.177

Abstract

Abstract:

BACKGROUND: Spheroid culture of mesenchymal stem cells in bioreactors is an in vitro culture method to maintain their stemness properties and allow for large-scale expansion. Clarifying its effects on the immunoregulation effect of stem cells is beneficial for their clinical application.

OBJECTIVE: To investigate the effects of spheroid culture in a rotating bioreactor on the secretion of inflammatory factors by human placenta-derived mesenchymal stem cells.
METHODS: Placenta-derived mesenchymal stem cells isolated from human placenta tissue were cultured in two-dimensional culture or a rotating bioreactor culture. Cell morphology and proliferation ability were observed using inverted phase contrast microscopy, immunohistochemical staining, and CCK-8 assay. The gene expression and protein secretion of several inflammatory factors were detected by RT-qPCR and flow immunofluorescence assay.
RESULTS AND CONCLUSION: (1) Mesenchymal stem cells cultured in a rotating bioreactor aggregated into multicellular spheroids, which gradually increased in number and volume. (2) The hematoxylin-eosin staining results showed that the mesenchymal stem cells cultured in the rotating bioreactor for 4 days were evenly distributed and had normal morphology in the spheroids. (3) Immunohistochemical staining results revealed many mesenchymal stem cells with Ki-67 positive in the spheroids. (4) The CCK-8 assay results exhibited that the viability of mesenchymal stem cells derived from spheroid culture was significantly higher than that of cells cultured in two-dimensional culture. (5) The results of RT-qPCR and flow immunofluorescence assay demonstrated that the gene expression and protein secretion (interleukin 1β, interleukin-4, interleukin-6, interleukin-8, interleukin-10, interleukin-17, tumor necrosis factor α and interferon α) of inflammatory factors derived from mesenchymal stem cells cultured in the rotating bioreactor were significantly higher than those in two-dimensional culture. (6) Our results indicate that spheroid culture in a rotating bioreactor can significantly elevate the secretion ability of various inflammatory factors by human placenta-derived mesenchymal stem cells, and enhance the immunoregulatory effect of human placenta-derived mesenchymal stem cells.

Key words: placenta-derived mesenchymal stem cell, bioreactor, spheroid, inflammatory factor, immunoregulation

CLC Number:

Zhang Pingping, Liang Tingting, Fan Mingsong, Chen Li, Zhang Shichang. Effects of spheroid culture in a rotating bioreactor on secretion of inflammatory factors by human placenta-derived mesenchymal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2024, 28(19): 3012-3017.

Figures/Tables 5

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