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

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

Research output: Contribution to journalArticle

Abstract

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, CD105, 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.

Original languageEnglish
Pages (from-to)648-657
Number of pages10
JournalIn Vitro Cellular and Developmental Biology - Animal
Volume54
Issue number9
DOIs
Publication statusPublished - 01-10-2018

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Stem Cells
Epithelium
Organoids
Lung
Vacuoles
CD15 Antigens
Alveolar Epithelial Cells
Induced Pluripotent Stem Cells
Periodic Acid
Regenerative Medicine
Serum
Suspensions
Transcription Factors
Enzymes
Research
Genes

All Science Journal Classification (ASJC) codes

  • Developmental Biology
  • Cell Biology

Cite this

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title = "A basic study on self-reconstitution of alveolar epithelium-like cells by tissue stem cells in mouse lung",
abstract = "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, CD105, 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.",
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A basic study on self-reconstitution of alveolar epithelium-like cells by tissue stem cells in mouse lung. / Hayashi, Masamichi; Yamamoto, Naoki; Hiramatsu, Noriko; Isogai, Sumito; Gotoh, Yusuke; Goto, Yasuhiro; Kondo, Masashi; Imaizumi, Kazuyoshi.

In: In Vitro Cellular and Developmental Biology - Animal, Vol. 54, No. 9, 01.10.2018, p. 648-657.

Research output: Contribution to journalArticle

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AU - Hayashi, Masamichi

AU - Yamamoto, Naoki

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