Oligopotent Mesenchymal Stem Cell‐Like Clone Becomes Multinucleated Following Phorbol Ester, TPA Stimulation

Abstract

We established a mesenchymal stem cell clone, 5F9A, from rat bone marrow substrate adherent cells by repeated limiting dilutions. The cells have a fibroblastic shape and form intimate contacts with adjacent cells with interdigitations and junctions similar to adherence and tight junctions in a semi-confluent culture. Analysis of the phenotypes of these cells by RT-PCR and FACS demonstrated that they resembled mesenchymal stem cells, and the cells could differentiate into adiopocytes and osteoblasts under appropriate conditions in vitro showing their oligopotency. Furthermore, the cells were induced to become multinuclear cells by TPA (12-o-tetradecanoylphorbol 13-acetate) stimulation.

Inhibition of Rho-associated kinases disturbs the collective cell migration of stratified TE-10 cells

“…Transmission electron microscopy images were captured as described previously [ 58 ]. The cells were fixed with 1.25 % glutaraldehyde and 1 % paraformaldehyde in 0.1 M phosphate buffer (pH 7.4), post-fixed with 1 % osmium tetroxide in the buffer, dehydrated with a graded series of ethanol, substituted by propylene oxide, and embedded in Epon 812.…”

Plasmodium induced by SU6656, an Src family kinase inhibitor, is accompanied by a contractile ring defect

“…9,10 They are induced to become multinucleated in the presence of TPA, a tumor promoter and a PKC activator. 9,10 On searching for other substances that have the ability to induce multinucleation in 5F9A cells, we identified SU6656, one of the potent SFK inhibitors (Figure 1a). During 3 days of incubation with SU6656, the cytoplasm of 5F9A cells increased in size, and in many cells, more than one nucleus was observed.…”

“…5F9A cells are mesenchymal stem cell‐like clones established from rat bone marrow adherent cells by repeated limiting dilutions . They are induced to become multinucleated in the presence of TPA, a tumor promoter and a PKC activator . On searching for other substances that have the ability to induce multinucleation in 5F9A cells, we identified SU6656, one of the potent SFK inhibitors (Figure a).…”

“…We reported that oligopotent mesenchymal stem cell‐like clones, 5F9A, changed into multinucleated cells by stimulation with phorbol ester, 12‐ O ‐tetradecanoylphorbol 13‐acetate (TPA), and this was mediated by regression of the cleavage furrow after slight ingression along the contractile ring, indicating the plasmodium mechanism . TPA induced about 10% of the cells to become multinucleated, although we could not determine what kind of multinuclear cells they differentiated into …”

“…9,10 TPA induced about 10% of the cells to become multinucleated, although we could not determine what kind of multinuclear cells they differentiated into. 9,10 In efforts to search for a potent inducer of multinucleation other than phorbol ester, we found that one Src family kinase (SFK) inhibitor, SU6656, induced the multinucleation of 5F9A mesenchymal stem cell-like clone at a higher rate than TPA.…”