Conversion of Goat Fibroblasts into Lineage-Specific Cells Using a Direct Reprogramming Strategy

Area of Science:

• Reproductive Biology
• Stem Cell Biology
• Cellular Reprogramming

Background:

• Maintaining undifferentiated goat embryonic stem- and induced pluripotent stem-like cells in vitro proved challenging.
• Direct reprogramming offers an alternative strategy for generating lineage-specific cells from fibroblasts.

Purpose of the Study:

• To explore direct reprogramming of goat fibroblasts into lineage-specific cells.
• To generate oocyte-like cells (OLCs) from reprogrammed goat fibroblasts.

Main Methods:

• Ectopic expression of 'Yamanaka factors' in fibroblasts to induce a metastable state.
• Culturing metastable cells in lineage-specific media for 1-2 weeks to generate cardiomyocyte-like and neurocyte-like cells.
• Culturing metastable cells in retinoic acid (RA) and bovine follicular fluid (bFF) for 2-3 weeks to generate oocyte-like cells (OLCs).

Main Results:

• Successfully generated cardiomyocyte-like and neurocyte-like cells, confirmed by quantitative RT-PCR and immunocytochemical staining.
• Generated OLCs resembling goat cumulus-oocyte complexes, which developed further and expressed germ cell-specific markers.
• Found higher induction efficiency and larger OLC size using bFF compared to RA treatment.

Conclusions:

• Direct reprogramming of goat fibroblasts is a viable strategy for generating lineage-specific cells, including OLCs.
• This method bypasses the difficulties associated with maintaining pluripotent stem cells in vitro.
• The findings facilitate stem cell research and potential applications in domestic animals.