Int J Stem Cells.  2020 Mar;13(1):104-115. 10.15283/ijsc19098.

Quantitative Tracking Tumor Suppression Efficiency of Human Umbilical Cord-Derived Mesenchymal Stem Cells by Bioluminescence Imaging in Mice Hepatoma Model

• Liu J
• Shi Y
• Han J
• Zhang Y
• Cao Z
• Cheng J

• KMID: 2500570
• DOI: http://doi.org/10.15283/ijsc19098

Abstract

Background and Objectives
Tracking of the tumor progression by MSCs-based therapy is being increasingly important in evaluating relative therapy effectively. Herein, Bioluminescence imaging (BLI) technology was used to dynamically and quantitatively track the hepatocellular carcinoma suppressive effects by human umbilical cord mesenchymal stem cells (UC-MSCs).
Methods and Results
The stem cells present typical phenotypic characteristics and differentiation ability by morphology and flow cytometry analysis of marker expression. Then, the growth inhibition effect of conditioned medium and UC-MSC on H7402 cells was studied. It is found both the conditioned medium and UC-MSC can effectively decrease the proliferation of H7402 cells compared with the control group. Meanwhile, the relative migration of UC-MSC to H7402 is also increased through the transwell migration assay. In addition, a mice hepatoma tumor model was built by H7402 cells which can express a pLenti-6.3/DEST-CMV-luciferase 2-mKate2 gene. The effect of stem cells on growth inhibition of tumor in a mice transplantation model was dynamically monitored by bioluminescence imaging within 5 weeks. It has shown the bioluminescence signal intensity of the tumor model was significantly higher than that of the UC-MSC co-acting tumor model, indicating that the inhibition of UC-MSC on liver cancer resulted in low expression of bioluminescent signals.
Conclusions
The microenvironment of UC-MSCs can effectively inhibit the growth of liver cancer cells, and this therapeutic effect can be dynamically and quantitatively monitored in vivo by BLI. This is of great significance for the imaging research and application of stem cells in anticancer therapy.

Keyword

Mesenchymal stem cells; Bioluminescent imaging; Hepatocellular carcinoma; Cancer therapy; Tumor environment

Figure

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  Fig. 1 (a) The cell morphology of UC-MSCs was observed under a light microscope (magnification, ×100). (b∼d) Representative images of osteocyte, adipocyte, and chondrocyte differentiation of UC-MSCs cultured in the differentiation media. The cells were analyzed using cytochemical staining with Alizarin Red, Oil red O, and Alcian Blue respectively. The experiment shown is representative of three performed (magnification, ×200).

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  Fig. 2 (a, b) Optical image and FACS of H7402 cells. (c, d) High-level expression of mKate2 in H7402 cells was demonstrated by fluorescence microscope and FACS. (e, f) In vitro imaging analysis of stably transduced H7402 cells shows a strong correlation between cell numbers and reporter gene activity. Scale bar represents 100 μm.

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  Fig. 3 (a) CCK-8 assay of H7402 cells with different concentration of UC-MSCs-CM for 96 h. Values are means±S.E. of three separate experiments performed (p<0.001). (b) The biological effects of different concentrations of UC-MSCs conditioned media on H7402 cell apoptosis.

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  Fig. 4 Migration ability of UC-MSCs to H7402 cells was examined using a transwell assay under different conditions. (a) Fresh 1640, (b) H7402 cells, (c) H7402-CM. (d) Average number of cells migrated in the transwell assay in different conditions. The results were expressed as the mean±S.D. in five different fields from three independent experiments (*p<0.05 and **p<0.01).

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  Fig. 5 Tumor growth monitored by bioluminescence imaging (BLI). (a) The bioluminescence images of the tumor-bearing mice (n=5) from the groups implanted with H7402 cells alone or (b) H7402 cells co-implanted with UC-MSCs in 35 days. (c) Imaging of the tumor-bearing mice from the groups implanted with H7402 cells alone or (d) H7402 cells co-implanted with UC-MSCs at 35 days post implantation. (e) Quantitative analysis of bioluminescence imaging signal.

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  Fig. 6 (a) Histopathological images of liver tumor tissues in experimental mice. HE and MVD staining of the tumor and the tumor treated with UC-MSCs. (b) The data are presented as the mean±standard deviation (SD).

Reference

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