Isolation and morphological characterization of equine mesenchymal stem cells from harvested adipose tissue and bone marrow and stably transfected with green fluorescent protein.

This research investigated the structure of mesenchymal stem cells (MSCs) gathered from the adipose tissue and bone marrow of horses. The collected cells were ultimately marked with a fluorescence protein for ease of study and comparison. The characteristics of those genetically marked cells were then studied and compared to their unmarked counterparts.

Methodology

• The study collected mesenchymal stem cells (MSCs) from the adipose tissue and bone marrow of 6 adult female Hispano-Bretón horses.
• The researchers then cultivated these harvested cells and proceeded to insert green fluorescent protein into their genetic makeup, a technique known as transfection.
• The identity or type of these cells, often referred to as the immunophenotypes, were determined using two specific antibodies, anti-CD90 and anti-CD105.
• Once the successful genetic marking (transfection) was confirmed, the researchers used electron microscopy to study the specifics of the cell structures in both transfected and non-transfected MSCs from adipose tissues and bone marrow.

Results

• The techniques used for transfecting cells and isolating those that had been successfully transfected showed favorable results.
• The study found that the rate of successful transfection was 5% for MSCs from adipose tissues and slightly lower at 4% for those from bone marrow.
• According to their morphological and immunochemical characteristics, the marked and unmarked cells were found to be largely similar.
• The expression of CD90 was found to be slightly higher in transfected cells compared to the unmarked adipose tissue MSCs, whereas the CD105 expression was lower in transfected cells compared to the unmarked bone marrow MSCs.
• Additionally, transfected cells originating from bone marrow exhibited noticeable differences in the organelles, particularly in the rough endoplasmic reticulum which is an organelle involved in protein synthesis and the mitochondria, which is responsible for energy production, compared to other cell types studied.

Conclusions

• The study’s findings contribute valuable information to the understanding of the characteristics of equine MSCs sourced from adipose tissue and bone marrow, as well as those of genetically marked and unmarked cells.
• This research can serve as a valuable reference for subsequent studies focusing on the morphological characteristics of equine MSCs.