Multilineage potential of side population cells from human amnion mesenchymal layer.
Side population (SP) cells were isolated by FACS from a human amnion mesenchymal cell (AMC) layer soon after enzyme treatment. The yield of SP cells from AMC layer (AMC-SP cells) was about 0.1-0.2%. AMC-SP cells grew well with cell doublings of 40-80 days of culture. FACS profiles and immunocytostaining showed that AMC-SP cells were composed of two different cells immunologically: HLA I(-)/II(-) and HLA P/II(-). Oct-3/4 was detected in the nucleus of AMC-SP cells, when the culture was examined at the third, sixth, and 10th passages. RT-PCR showed that AMC-SP cells expressed the Oct-4, Sox-2, and Rex-1 genes. Immunocytochemistry revealed that all AMC-SP cells were vimentin+, CK19+, and nestin+. In addition, flow cytometry analysis showed that SP cells had high expression of CD13, CD29, CD44, CD46, CD49b, CD49c, CD49e, CD59, CD140a, and CD166 but low expression of CD 49d, and CD51. No evidence of expression was obtained for CD34, CD45, CD49a, CD56, CD90, CD105, CD106, CD117, CD133, CD271, or Flk-1. Upon appropriate differentiation protocols, AMC-SP cells differentiated to several cell lineages such as neuroectodermal, osteogenic, chondrogenic, and adipogenic cells. These results indicate that AMC-SP cells have multilineage potential to several cell lineages with unique immunological characteristics such as HLA I(-)/II(-) or HLA I+/II(-). AMC-SP cells should be of considerable value for regenerative medicine because they do not induce acute rejection after allotransplantation, they do not cause ethical issues, and there is no limit of supply.
Department of Regenerative Medicine, School of Allied Health Sciences, Kitasato University, Nishi-hashimoto, Kanagawa, Japan.
M
M
Kobayashi
T
T
Yakuwa
K
K
Sasaki
K
K
Sato
A
A
Kikuchi
I
I
Kamo
Y
Y
Yokoyama
N
N
Sakuragawa
D017667:Adipocytes / Q000166:cytology / Q000378:metabolism; D000650:Amnion / Q000166:cytology* / Q000378:metabolism; D018826:CD13 Antigens / Q000032:analysis; D002454:Cell Differentiation; D019070:Cell Lineage; D019902:Chondrocytes / Q000166:cytology / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics; D005434:Flow Cytometry; D024002:HMGB Proteins / Q000235:genetics; D006801:Humans; D007150:Immunohistochemistry; D019012:Integrin beta1 / Q000032:analysis; D007381:Intermediate Filament Proteins / Q000032:analysis; D053539:Keratin-19 / Q000032:analysis; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000032:analysis; D064231:Nestin; D050814:Octamer Transcription Factor-3 / Q000235:genetics; D010011:Osteocytes / Q000166:cytology / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D055748:SOXB1 Transcription Factors; D014157:Transcription Factors / Q000235:genetics; D014746:Vimentin / Q000032:analysis
D004268:DNA-Binding Proteins; D024002:HMGB Proteins; D019012:Integrin beta1; D007381:Intermediate Filament Proteins; D053539:Keratin-19; C577484:NES protein, human; D009419:Nerve Tissue Proteins; D064231:Nestin; D050814:Octamer Transcription Factor-3; C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors; D014157:Transcription Factors; D014746:Vimentin; D018826:CD13 Antigens