Effects of cell heterogeneity on production of polypeptide growth factors and mesoderm-inducing activity by Xenopus laevis XTC cells.
The Xenopus laevis XTC cell line has been analyzed for the production of polypeptide growth factors and mesoderm-inducing activity. By the use of specific biological assays, it is shown that XTC cells produce a growth factor functionally related to the platelet-derived growth factor (PDGF) and two transforming growth factor (TGF) beta-like activities. Mesoderm-inducing activity, as measured on X. laevis ectodermal explants from stage 10 embryos, was found to coelute on a Bio-Gel P-100 column with one of the TGF beta-like activities at an apparent molecular weight of 6-10 kDa. Analysis of the DNA content from XTC cells by flow cytometry demonstrated that the cell line is heterogeneous and consists of both tetraploid and diploid cells. Cloning of the XTC cells and selecting single-cell colonies on the basis of their ability to grow in soft agar resulted in the isolation of several homogeneous, morphologically different clonal derivatives. Analysis of conditioned medium from these clonal derivatives showed that only one of them, the only diploid line among six investigated, produced a strong heat- and acid-stable mesoderm-inducing activity that induced notochord and muscle formation in stage 10 X. laevis ectodermal explants. The relation between this activity and a recently described TGF beta-like mesoderm-inducing factor obtained from XTC-conditioned medium will be discussed. In conclusion, a clonal cell line derived from X. laevis XTC cells which provides a good source for further characterization of mesoderm-inducing factors has been established.
Snoek
G T
GT
Hubrecht Laboratory, Netherlands Institute for Developmental Biology, Utrecht.
Koster
C H
CH
de Laat
S W
SW
Heideveld
M
M
Durston
A J
AJ
van Zoelen
E J
EJ
D000818:Animals; D002460:Cell Line; D002850:Chromatography, Gel; D002999:Clone Cells; D004247:DNA / Q000032:analysis; D004625:Embryo, Nonmammalian / Q000502:physiology; D005434:Flow Cytometry; D006133:Growth Substances / Q000096:biosynthesis* / Q000302:isolation & purification; D008648:Mesoderm / Q000166:cytology / Q000502:physiology*; D008970:Molecular Weight; D010452:Peptide Biosynthesis; D010455:Peptides / Q000302:isolation & purification; D014982:Xenopus laevis
D006133:Growth Substances; D010455:Peptides; D004247:DNA