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ORIGINAL RESEARCH ARTICLE

Culture Supernatant of Lactobacillus acidophilus Stimulates Proliferation of Embryonic Cells

W. I. Li^*,1, Benjamin G. Brackett^* and Jaroslava Halper

^* Department of Physiology and Pharmacology; and Department of Pathology, College of Veterinary Medicine, The University of Georgia, Athens, Georgia 30602

To whom requests for reprints should be addressed at ¹ Department of Physiology and Pharmacology, College of Veterinary Medicine, The University of Georgia, Athens, GA 30602. E-mail: oliverli{at}vet.uga.edu

Our previous report showed that supernatants of Lactobacillus acidophilus (LS) cultures possessed chemotactic and angiogenic properties. Specifically, LS stimulated gene expression and the secretion of tumor necrosis factor- (TNF-), the proliferation of immune cells in vitro, and blood vessel formation. Chemotaxis and proliferation of inflammatory cells in vivo were also stimulated by LS. In the current study, we hypothesized that LS stimulates the growth and development of other rapidly dividing cells, including embryonic cells. The stimulatory effects of LS on a neuroblastoma cell line (Neuro-2a), chicken embryos, and bovine embryos were examined. The addition of LS to Neuro-2a cultures caused a proliferation of cells in a concentration-dependent manner. Pretreatment of LS at 56°C for 30 mins did not affect its stimulatory activity. The administration of LS to the chorioallantoic membrane (CAM) of chicken-embryonated eggs for 1–2 days resulted in extensive thickening of the membrane. The thickening was due to the influx and proliferation of fibroblasts and inflammatory cells, the accumulation of loose connective tissue composed primarily of mucopolysaccharides, and/or the formation of blood vessels. Stimulatory effects of LS on bovine embryos were also observed. The treatment with LS significantly promoted the development of zygotes to the four-cell stage and from the four-cell stage to blastocysts. These results have confirmed our hypothesis that LS exerts a stimulatory effect on the cells of embryonic stages including neuroblastoma cells, the CAM of chicken embryos, and bovine embryos from zygotes to blastocysts.

Key Words: Lactobacillus supernatant • Neuro-2a cells • chicken chorioallantoic membrane • bovine embryos