Tissue Eng Regen Med.  2018 Oct;15(5):649-659. 10.1007//s13770-018-0132-z.

Characterization of Human Fetal Cartilage Progenitor Cells During Long-Term Expansion in a Xeno-Free Medium

• Kim HR
• Kim J
• Park SR
• Min BH
• Choi BH

• KMID: 2422678
• DOI: http://doi.org/10.1007//s13770-018-0132-z

Abstract

BACKGROUND
Stem cell therapy requires a serum-free and/or chemically-defined medium for commercialization, but it is difficult to find one that supports long-term expansion of cells without compromising their stemness, particularly for novel stem cells.
METHODS
In this study, we tested the efficiency of StemPro® MSC SFM Xeno Free (SFM-XF), a serum-free medium, for the long-term expansion of human fetal cartilage-derived progenitor cells (hFCPCs) from three donors in comparison to that of the conventional α-Modified Eagle's Medium (α-MEM) supplemented with 10% fetal bovine serum (FBS).
RESULTS
We found that SFM-XF supported the expansion of hFCPCs for up to 28-30 passages without significant changes in the doubling time, while α-MEM with 10% FBS showed a rapid increase in doubling time at 10-18 passages depending on the donor. Senescence of hFCPCs was not observed until passage 10 in both media but was induced in approximately 15 and 25% of cells at passage 20 in SFM-XF and α-MEM with 10% FBS, respectively. The colony forming ability of hFCPCs in SFX-XF was also comparable to that in α-MEM with 10% FBS. hFCPCs expressed pluripotency genes like Oct-4, Sox-2, Nanog, SCF, and SSEA4 at passage 20 and 31 in SFM-XF; however, this was not observed when cells were cultured in α-MEM with 10% FBS. The ability of hFCPCs to differentiate into three mesodermal lineages decreased gradually in both media but it was less significant in SFM-XF. Finally we found no chromosomal abnormality after long-term culture of hFCPCs until passage 17 by karyotype analysis.
CONCLUSION
These results suggest that SFM-XF supports the long-term expansion of hFCPCs without significant phenotypic and chromosomal changes. This study have also shown that hFCPCs can be mass-produced in vitro, proving their commercial value as a novel source for developing cell therapies.

Keyword

Human fetal cartilage progenitor cells; Serum-free medium; Cell therapy; Pluripotency

MeSH Terms

Aging
Cartilage*
Cell- and Tissue-Based Therapy
Chromosome Aberrations
Humans*
In Vitro Techniques
Karyotype
Mesoderm
Stem Cells*
Tissue Donors