Establishment of a clonal human mesenchymal cell line that retains multilineage differentiation capacity from a spinal hamartoma.
We isolated a single-cell-derived cell line from a spinal hamartoma, a which occurred in a newborn boy and was associated with a rudimentary limb. The maternal cells (HHC-7) differentiated into osteoblasts, chondrocytes, adipocytes, and skeletal muscles when they were cultured in differentiation-inducing media specific to each mesenchymal cell. We isolated a single-cell-derived clonal cell line (Clone K) after transfection with SV40 T antigen. These cells expressed CD73 and CD117, while being negative for expression of CD45. Clone K cells cultured in an osteogenic differentiation medium increased ALP activity and expressed mRNAs for Runx2 and osteocalcin. Treatment with rhBMP-2 induced Clone K cells to differentiate into both osteoblasts and chondrocytes. These cells expressed mRNAs for Sox9 and aggrecan in addition to osteogenic markers. Culture in an adipogenic differentiation medium induced Clone K cells to differentiation into adipocytes, which expressed mRNAs for PPARgamma2 and a2P. Clone K cells cultured in a serum-depleted medium generated desmin-positive cells and expressed MyoD1 mRNA. Clone K cells exhibited numerous alpha-smooth muscle actin-positive cells; however, treatment with rhBMP-2 decreased their number. Clone K cells, transplanted with a carrier containing rhBMP-2 into the muscles of SCID mice, generated ectopic endochondral bone formation. In these tissues, several osteoblasts and chondrocytes expressed SV40 T antigen, indicating their Clone K cell origin. Thus, Clone K cells are useful tools for analyzing the characteristics of human multipotential mesenchymal progenitors.
Division of Orthopaedic Pathomechanism, Department of Developmental and Reconstructive Medicine, Nagaski University Graduate School of Biomedical Sciences, Nagaski, Japan.
Y
Y
Doiguchi
T
T
Tsukazaki
T
T
Tomonaga
M
M
Nobuta
S
S
Fujita
T
T
Hayashi
K
K
Nagai
T
T
Matsumoto
H
H
Shindo
A
A
Yamaguchi
D017667:Adipocytes / Q000166:cytology; D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D019902:Chondrocytes / Q000166:cytology; D002999:Clone Cells; D006222:Hamartoma / Q000378:metabolism / Q000473:pathology* / Q000601:surgery; D006801:Humans; D007231:Infant, Newborn; D008297:Male; D008648:Mesoderm / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation; D051379:Mice; D016513:Mice, SCID; D018482:Muscle, Skeletal / Q000166:cytology; D010006:Osteoblasts / Q000166:cytology; D013122:Spinal Diseases / Q000378:metabolism / Q000473:pathology* / Q000601:surgery; D014162:Transfection