Isolation and Culture of Bovine Embryonic Stem Cells

Abstract

Isolation and culture of primary embryonic stem (ES) cell colonies are the first critical step towards establishment of stable ES cell lines. Here we introduce a novel method designated as "Separate and Seed" that contributes remarkably to efficient derivation of bovine primary ES cell colonies from blastocysts. The bovine ES cell colonies can self-renew to passage 10 with the growth factors bFGF and BIO. The bovine ES cells exhibit morphology typical of ES cells and express pluripotent molecular markers including Oct4, Nanog, SSEA1, SSEA4, and alkaline phosphatase (AP). These pluripotent markers may be used for the characterization of authentic bovine ES cell lines. Although continued efforts are required for improving long-term culture of bovine ES cells, this novel "Separate and Seed" method plus the growth factors bFGF and BIO provides an initial effective step that may eventually lead to the derivation of authentic bovine ES cells.

Cell Sources for Cultivated Meat: Applications and Considerations throughout the Production Workflow

“…However, the base pair sequence and identity of these factors may vary depending on the species. For example, bovine ESC [ 33 ] and iPSC [ 39 ] markers have been determined to express Oct4 and Nanog, but also Stage-specific embryonic antigen (SSEA) 1, SSEA3, and SSEA4, which are not expressed in human pluripotent stem cells. Identifying species differences in pluripotency factors may represent an important first step in developing pluripotent cell lines from livestock species.…”

“…In the second part, we will discuss the subsequent components of the workflow with special focus being paid to cellular considerations during these production steps (Table 2). Bovine [18] Galline [19] Ovine [20] Piscine [21] Porcine [22] Mesenchymal stem/stromal cells (MSCs) Bovine [33] Galline [34] Ovine [35] Piscine [36,37] Porcine [38] Induced pluripotent stem cells (iPSCs) Bovine [39] Galline [40,41] Ovine [42] Porcine [43] Table 2. Overview of advancements and current limitations in the cultivated meat production workflow.…”

The Stem Cell Niche as the Key to Early Cancer Development

“…Currently several components required for stem cells growth, for example, have been identified ( Figure 2). Among them basic fibroblast growth factor (bFGF) has been shown to be essential for self-renewal in embryonic stem cells [20,21]. Three other requirements are (i) feeder cells, conditioned medium, or cytokines, such as TGF [22,23] or Wnt3a [24]; (ii) matrix; and (iii) FBS or serum replacement [25,26].…”