Identification of an aberrant cell line among human adipose tissue-derived stem cell isolates.
Adipose tissue-derived stem cells (ADSC) are isolated from the stromal vascular fraction (SVF) of adipose tissue and considered an excellent cell source for regenerative medicine. During the isolation and propagation of several human ADSC cell lines, we observed the emergence of an unusual cell line designated HADSC-6. Although initially fibroblast-like as typical ADSC are, HADSC-6 cells became homogeneously cuboid in shape, had very little cytoplasm, and formed aggregates with capsule-like boundary. Proliferation assay showed that HADSC-6 grew much faster than typical HADSC cell lines, such as HADSC-20. Immunocytochemistry showed that HADSC-6 did not express endothelial markers CD31 and vWF, and matrigel tube formation assay showed that it was unable to form endothelial-like tube structures. However, LDL uptake, a reliable endothelial marker, was positively identified. Chromosomal analysis showed that HADSC-6 cells were hypertriploid, and soft agar colony formation assay showed that they were able to proliferate and form large colonies in an anchorage-independent manner. However, tumorigenicity test showed that HADSC-6 was unable to form tumors in athymic mice. RT-PCR analysis showed that both HADSC-6 and HADSC-20 expressed VEGF-A, VEGF-B, VEGF-D, and VEGFR1 but not VEGFR2 or VEGFR3. VEGF-C, however, was expressed at a high level in HADSC-20 but undetectable in HADSC-6. In the IGF system, IGF-1 was abundantly expressed in HADSC-20 but marginally detectable in HADSC-6, and IGF-1R was abundantly expressed in HADSC-6 but not detectable in HADSC-20. In the FGF system, bFGF was abundantly expressed in HADSC-20 but marginally detectable in HADSC-6, and FGFR1 was abundantly expressed in both. Taken together, these results suggested that HADSC-6 cells were spontaneously transformed from the endothelium; therefore, they were further compared to previously published data of four naturally occurring human angiosarcoma cell lines. The results showed that the established angiosarcoma cell lines exhibit considerable variations among themselves and HADSC-6 displayed most of these variable characteristics.
Ning
Hongxiu
H
Knuppe Molecular Urology Laboratory, Department of Urology, School of Medicine, University of California, San Francisco, CA 94143-0738, USA.
Liu
Gang
G
Lin
Guiting
G
Garcia
Maurice
M
Li
Long-Cheng
LC
Lue
Tom F
TF
Lin
Ching-Shwun
CS
D000273:Adipose Tissue / Q000166:cytology* / Q000378:metabolism; D000818:Animals; D018929:Cell Culture Techniques; D002460:Cell Line; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000473:pathology / Q000502:physiology*