Evaluation of the Chondrogenic Differentiation Capacity of Rabbit Umbilical Cord Stem Cells
Authors: Kruse Lauren , Rachakatla Raja S. 
Published: April 2020

The long‐term objective of this study is to evaluate regenerative therapies for injured tissues and to improve treatment options. Osteoarthritis is one of the most common causes of pain and disability, affecting approximately 30 million people in the United States alone and increasing. Osteoarthritis is a complicated disease that involves changes in the subchondral bone, articular cartilage degeneration, and surrounding soft tissue damage. Despite the global increase in osteoarthritis, conventional pharmacotherapies that can repair the structure and function are limited. Surgical interventions have some success, however, they come with risks of failure and infection as well as the costs of hospital care and rehabilitation. The lack of conventional treatment and the high costs of surgery has driven research towards cell therapy and tissue engineering approaches focused particularly on stem cells as an option for treating osteoarthritis. Umbilical cord tissue is considered a promising source of mesenchymal stem cells that could possibly be used for clinical applications that are free from ethical complications and are easy to isolate. Umbilical cord mesenchymal stem cells have a profile of gene expression similar to embryonic stem cells and they have faster self‐renewal.

Aim

In this study, our aim was to evaluate the potential of rabbit umbilical cord stem cells differentiation into chondrocytes for its possible use in cartilage repair therapy for osteoarthritis in the future.

Methods

Rabbit umbilical cord stem cells were grown in T75 flasks in DMEM with 10% fetal bovine serum and 1% antibiotic. The goal was to have approximately 15,000–20,000 cells per well in a 6 well plate or about 7,500–10,000 cells per well in a 12 well plate. We divided the wells into a control group (DMEM media) and experimental group (STEMPRO Chondrogenesis differentiation media) and incubated for 21–30 days with media changed every 5–7 days. Pictures were taken each day to evaluate morphological changes of the cells. After 3–4 weeks, chondrogenic differentiation was analyzed via immunocytochemistry for Collagen II monoclonal antibody, and alcian blue staining for proteoglycans. Bright field images were taken with Evos XL core and fluorescent images were taken with EVOS FL Color imaging systems. All experiments were done in triplicate and were repeated thrice.

Results

Our results show that rabbit umbilical cord stem cells in a monolayer cell culture differentiated easily into chondrocytes after 3–4 weeks of chondrogenic induction assessed by immunocytochemistry and alcian blue staining. This is compared to cells without chondrogenesis differentiation media, that were negative for both collagen type II and proteoglycans.

Conclusion

This study demonstrates that rabbit umbilical cord stem cells, a model for human umbilical cord mesenchymal stem cells, have the potential to differentiate into chondrocytes effectively and could possibly have potential for cartilage tissue engineering.

Support or Funding Information

Arkansas College of Osteopathic Medicine Research Grant