PLURIPOTENT CELL-LINES DERIVED FROM COMMON MARMOSET (CALLITHRIX-JACCHUS) BLASTOCYSTS

We report the derivation of eight pluripotent cell lines from common m armoset (Callithrix jacchus) blastocysts. These cell lines are positiv e for a series of markers (alkaline phosphatase, SSEA3, SSEA-4, TRA-1- 60, and TRA-1-81) that characterize undifferentiated human embryonal c arcinoma cells and rhesus embryonic stem cells. All eight cell lines h ad a modal chromosome number of 46; seven cell lines were XX and one w as XY. Two cell lines (Cj11 and Cj62) were cultured continuously for o ver a year and remained undifferentiated and euploid. In the absence o f fibroblast feeder layers, these cell lines differentiated to multipl e cell types, even in the presence of leukemia inhibiting factor. Diff erentiated cells secreted bioactive CG into the culture medium and exp ressed alpha-CG, beta-CC, and alpha-fetoprotein mRNA, indicating troph oblast and endoderm differentiation. Bioactive CG secretion in differe ntiating cells was increased substantially in the presence of GnRH ago nist D-Trp(6)-Pro(9)-NHEt. When grown at high densities, these cells f ormed embryoid bodies with a close resemblance to early postimplantati on embryos, including the formation of a yolk sac, amnion, and an embr yonic disc with an early primitive streak. These results make these pl uripotent cells strong candidates for marmoset embryonic stem cells.