title: Isolation of canine mesenchymal stem cells from amniotic fluid and differentiation into hepatocyte-like cells.
abstract: Recent findings have demonstrated that amniotic fluid cells are an interesting and potential source of mesenchymal stem cells (MSCs). In this study, we isolated MSCs from canine amniotic fluid and then characterized their multilineage differentiation ability. Canine amniotic fluid stem (cAFS) cells at passage 5 had a fibroblast-like morphology instead of forming colonies and were positive for pluripotent stem cell markers such as OCT4, NANOG, and SOX2. Flow cytometry analysis showed the expression of MSC surface markers CD44, CD29, and CD90 on the cAFS cells. In addition, these cells were cultured under conditions favorable for adipogenic, chondrogenic, and osteogenic induction. The results of this experiment confirmed the mesenchymal nature of cAFS cells and their multipotent potential. Interestingly, although the cells exhibited a fibroblast-like morphology after hepatogenic induction, reverse transcription-polymerase chain reaction revealed that the expression of several hepatic genes, such as albumin, tyrosine aminotransferase, and alpha-1 antiproteinase, increased following maturation and differentiation. These findings indicated that cAFS cells have functional properties similar to those of hepatocytes. Taken together, the results of our study demonstrated that cAFS cells with mesenchymal characteristics can be successfully isolated from canine amniotic fluid and possess functional properties characteristic of hepatocytes. The findings of our work suggest that cAFS cells have the potential to be a resource for cell-based therapies in a canine model of hepatic disease.
authors.affiliation: Department of Animal Science and Biotechnology, College of Agriculture and Life Science, Chungnam National University, Daejeon, 305-764, Republic of Korea.
authors.forename: Seon-A
authors.initials: SA
authors.lastname: Choi
authors.forename: Hoon-Sung
authors.initials: HS
authors.lastname: Choi
authors.forename: Keun Jung
authors.initials: KJ
authors.lastname: Kim
authors.forename: Dong-Soo
authors.initials: DS
authors.lastname: Lee
authors.forename: Ji Hey
authors.initials: JH
authors.lastname: Lee
authors.forename: Jie Yeun
authors.initials: JY
authors.lastname: Park
authors.forename: Eun Young
authors.initials: EY
authors.lastname: Kim
authors.forename: Xiaoxia
authors.initials: X
authors.lastname: Li
authors.forename: Hyun-Yang
authors.initials: HY
authors.lastname: Oh
authors.forename: Dong-Seok
authors.initials: DS
authors.lastname: Lee
authors.forename: Min Kyu
authors.initials: MK
authors.lastname: Kim
mesh_terms: D000418:Albumins / Q000378:metabolism; D000653:Amniotic Fluid / Q000166:cytology*; D000704:Analysis of Variance; D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000502:physiology*; D017931:DNA Primers / Q000235:genetics; D004285:Dogs; D005434:Flow Cytometry; D022781:Hepatocytes / Q000166:cytology*; D018960:Hyaluronan Receptors / Q000378:metabolism; D007150:Immunohistochemistry; D019012:Integrin beta1 / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000502:physiology; D011480:Protease Inhibitors / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018800:Thy-1 Antigens / Q000378:metabolism; D014444:Tyrosine Transaminase / Q000378:metabolism
chemical_list: D000418:Albumins; D017931:DNA Primers; D018960:Hyaluronan Receptors; D019012:Integrin beta1; D011480:Protease Inhibitors; D018800:Thy-1 Antigens; D014444:Tyrosine Transaminase