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

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 Ò 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 a-Modified Eagle's Medium (a-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 a-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 a-MEM with 10% FBS, respectively. The colony forming ability of hFCPCs in SFX-XF was also comparable to that in a-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 a-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. Keywords Human fetal cartilage progenitor cells Á Serum-free medium Á Cell therapy Á Pluripotency Hwal Ran Kim and Jiyoung Kim have contributed equally to this study.

Bringing Safe and Standardized Cell Therapies to Industrialized Processing for Burns and Wounds

“…Optimization of this selection process, with major emphasis set on safety and consistency, has led to the identification and recognition of allogenic primary progenitor cell types as highly promising and efficient candidates for cell therapies (De Buys Table 2. Roessingh et al, 2006;Mirmalek-Sani et al, 2006;Grognuz et al, 2016;Kim et al, 2018). Medical needs in the domains of burns and chronic cutaneous wound management are difficult to meet, due to the complex nature of wound bed environments and the delicate process of coordinated responses governing wound closure.…”

Immunophenotype and Immune-Modulatory Activities of Human Fetal Cartilage-Derived Progenitor Cells

“…Although the availability of donor tissue is limited, a fetal cartilage yields more than 20-fold the number of cells than the same amount of adult cartilage. In addition, we have also shown that hFCPCs can be expanded for more than 30 passages without losing their proliferation ability in a xeno-free medium, which resulted in approximately 10 30 cells from 5 × 10 5 cells of initial culture 57 . Therefore, we believe hFCPCs have a competitive commercial value, but understand that actual commercialization requires further investigation on their characteristics and behavior during long-term expansion and development of means for mass production and quality control as well.…”

“…passages without losing their proliferation ability in a xenofree medium, which resulted in approximately 10 30 cells from 5 Â 10 5 cells of initial culture 57 . Therefore, we believe hFCPCs have a competitive commercial value, but understand that actual commercialization requires further investigation on their characteristics and behavior during long-term expansion and development of means for mass production and quality control as well.…”

Holistic Approach of Swiss Fetal Progenitor Cell Banking: Optimizing Safe and Sustainable Substrates for Regenerative Medicine and Biotechnology

“…Pragmatic optimization of cell source selection and processing is crucial within translational development and clinical implementation of cell therapies and related products. Iterative amelioration and successful application of standardized workflows have led to identify allogenic primary FPC sources as highly promising and efficient candidates for regenerative medicine (Hebda and Dohar, 1999;De Buys Roessingh et al, 2006;Mirmalek-Sani et al, 2006;Ferguson, 2007, 2008;Larijani et al, 2015;Grognuz et al, 2016b;Kim et al, 2018). Upon adequate isolation from fetal tissues (i.e., enzymatic or mechanical methods), culture-expansion and cryopreservation, progeny cells and derivatives present numerous advantages.…”