Journal Article

A basic study on self-reconstitution of alveolar epithelium-like cells by tissue stem cells in mouse lung

Yamamoto, Naoki, Goto, Yasuhiro, Hayashi, Masamichi, Kondo, Masashi, Gotoh, Yusuke, Imaizumi, Kazuyoshi, Hiramatsu, Noriko, Isogai, Sumito

2018

Overview

In recent research on regenerative medicine, three-dimensional (3D) tissue reconstruction using the induced pluripotent stem cell (iPS cell) differentiated cells has attracted attention. In this study, mouse lungs at 1.5,10, and 20 d old were subjected to enzyme treatment, and aggregates formed in serum-free suspension culture (3D-culture) were observed. The number of aggregates formed was the highest in 1.5 d. The cell aggregates in which the interior of the aggregate is filled and form small vacuoles and the organoid-like aggregates having a relatively large vacuole inside and forming the alveolar-like structure were observed. At 1.5 d, the formation ratio of the organoid-like aggregates was the highest and aggregate size was small at 20 d. For the cell aggregates derived from 1.5 d, positive cells of SSEA-1, CD29, CD90, CD 105, alveolar epithelial stem cell marker of SP-C, and Sca-1 were observed in the center. In the cell aggregates derived from 10 d, the expression level of 1.5 d each protein markers and OCT4 gene of transcription factor was decreased, and furthermore, markers were hardly observed in the organoid-like aggregates derived from 10 d. In addition, cells surrounding the vacuole of organoid-like aggregate obtained over 10 d differentiated into periodic acid-Schiff (PAS), podoplanin-positive cells. When the formed cell aggregates were dispersed, cell aggregates and organoid-like aggregates were reformed. Comparing 3D-culture and adhesion culture (2D-culture), SP-C expression of 10 d of cells was maintained. Expression of markers of undifferentiated markers and alveolar tissue stem cells decreased when cell aggregates were cultured with the addition of fetal bovine serum.

• Publisher: Springer Science & Business Media LLC,Springer US,Society for In Vitro Biology
• Subject: adhesion / Aggregates / Alveoli / Animal Genetics and Genomics / Animals / Antibodies / Biomarkers / Biomarkers - metabolism / Biomedical and Life Sciences / CD105 antigen / CD29 antigen / CD90 antigen / Cell Adhesion / cell aggregates / Cell Aggregation / CELL AND TISSUE MODELS / Cell Biology / Cell Culture / Cell Differentiation / Cell Shape / Cells, Cultured / Chronic obstructive pulmonary disease / Developmental Biology / enzymatic treatment / epithelial cells / Epithelium / fetal bovine serum / Gene expression / Gene Expression Regulation / genes / Image reconstruction / in vitro studies / Life Sciences / Lungs / medicine / mice / Mice, Inbred C57BL / Oct-4 protein / Organoids / Penicillin / Pluripotency / Pulmonary Alveoli - cytology / Regenerative medicine / Respiratory Mucosa - cytology / Staining and Labeling / Stains & staining / Stem cell transplantation / Stem Cells / Stem Cells - cytology / Stem Cells - metabolism / Surfactants / Suspension culture / Tissue engineering / transcription factors / Transplants & implants / Vacuoles