J Korean Assoc Pediatr Surg.  2009 Jun;15(1):1-10.

Isolation and Identification of Respiratory Cells from Human Amniotic Fluid

Affiliations
  • 1Department of Surgery, Yonsei University College of Medicine, Seoul, Korea. jtoh@yuhs.ac
  • 2Department of Obstetrics and Gynecology, Yonsei University College of Medicine, Seoul, Korea.
  • 3The Research Institute for Transplantation, Yonsei University College of Medicine, Seoul, Korea.

Abstract

Recently, amniotic fluid has gained attention as one of the potential sources for cell therapy and tissue engineering because it has characteristics of multipotent stem cells. However, current knowledge about what types of cells are naturally found in amniotic fluid is still limited. In this study, we aimed to investigate whether human amniotic fluid contains cells that have characteristics of respiratory cells. Samples of human amniotic fluid (5 mL per sample) obtained from amniocenteses were cultured with small airway growth medium (SAGM). Cells were grown until the third passage and the presence of type II alveolar cells were characterized by inverted microscopy, immunofluorescence, and reverse transcription polymerase chain reaction (RT-PCR). On inverted microscopy, cultured cells showed typical polygonal and cobblestone-like epithelial morphology. The morphology of cells was not changed after selection and passing. Immunofluorescence analysis demonstrated that the isolated cells stained positive for surfactant protein C (SPC), specific marker for type II alveolar cells. Cells also stained positive for TTF-1 protein but negative for CD 31 and vimentin. RT-PCR analysis of cells showed expression of SPC mRNA. This study has demonstrated that respiratory cells can be isolated and identified from human amniotic fluid cultured in SAGM medium. Our results may provide the basis for further investigations of amniotic fluid.

Keyword

Amniotic fluid; Respiratory cell; Isolation; Human

MeSH Terms

Amniocentesis
Amniotic Fluid
Cells, Cultured
Female
Fluorescent Antibody Technique
Humans
Microscopy
Microscopy, Fluorescence
Multipotent Stem Cells
Polymerase Chain Reaction
Protein C
Reverse Transcription
RNA, Messenger
Tissue Engineering
Tissue Therapy
Vimentin
Protein C
RNA, Messenger
Vimentin
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