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An intra-amniotic injection of mesenchymal stem cells promotes lung maturity in a rat congenital diaphragmatic hernia model

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Abstract

Purpose

We aimed to evaluate the effect of human mesenchymal stem cells (hMSCs) on congenital diaphragmatic hernia (CDH) by intra-amniotic injection in a rat CDH model.

Methods

Nitrofen (100 mg) was administered to pregnant rats at E9.5. hMSCs (1.0 × 106) or PBS was injected into each amniotic cavity at E18, and fetuses were harvested at E21. The fetal lungs were classified into normal, CDH, and CDH-hMSCs groups. To determine the lung maturity, we assessed the alveolar histological structure by H&E and Weigert staining and the alveolar arteries by Elastica Van Gieson (EVG) staining. TTF-1, a marker of type II alveolar epithelial cells, was also evaluated by immunohistochemical staining and real-time reverse transcription polymerase chain reaction.

Results

The survival rate after intra-amniotic injection was 72.1%. The CDH-hMSCs group had significantly more alveoli and secondary septa than the CDH group (p < 0.05). The CDH-hMSCs group had larger air spaces and thinner alveolar walls than the CDH group (p < 0.05). The medial and adventitial thickness of the pulmonary artery in the CDH-hMSCs group were significantly better (p < 0.001), and there were significantly fewer TTF-1-positive cells than in the CDH group (p < 0.001).

Conclusion

These results suggest that intra-amniotic injection of hMSCs has therapeutic potential for CDH.

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Acknowledgements

This work was supported in part by Grant-in-Aid for Exploratory Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan (MEXT KAKENHI grant number 15K10926 [TF]). The English used in this manuscript was reviewed by Brian Quinn (Editor-in-Chief, Japan Medical Communication).

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Authors and Affiliations

  1. Department of Pediatric Surgery, Kyoto Prefectural University of Medicine, 465 Kawaramachi-Hirokoji, Kamigyo-ku, Kyoto, 602-8566, Japan

    Shohei Takayama, Kohei Sakai, Shigehisa Fumino, Taizo Furukawa & Tatsuro Tajiri

  2. Department of Immunology, Kyoto Prefectural University of Medicine, Kyoto, Japan

    Shohei Takayama, Tsunao Kishida & Osam Mazda

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  1. Shohei Takayama
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  2. Kohei Sakai
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Correspondence to Shohei Takayama.

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Takayama, S., Sakai, K., Fumino, S. et al. An intra-amniotic injection of mesenchymal stem cells promotes lung maturity in a rat congenital diaphragmatic hernia model. Pediatr Surg Int 35, 1353–1361 (2019). https://doi.org/10.1007/s00383-019-04561-7

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  • DOI: https://doi.org/10.1007/s00383-019-04561-7

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