TY - JOUR

T1 - Induction of Chondrogenic Differetiation of Human Dermal Fibroblasts Using Protein-Based Differentiation Media

AU - Petyukevich, V. P.

AU - Kovaleva, E. A.

AU - Bozhkova, S. A.

AU - Sopova, J. V.

AU - Leonova, E. I.

AU - Aleksandrova, S. A.

AU - Khotin, M. G.

AU - Bozhokin, M. S.

PY - 2025/8/19

Y1 - 2025/8/19

N2 - Abstract: Background: Existing methods of repairing articular cartilage used in medical practice are not fully effective, and therefore, the development of new approaches is a high priority for regenerative medicine. One promising approach is the use of tissue engineering techniques, which involve the use of biomedical cellular products containing biodegradable scaffolds and modified cells. Objective: This study aims to evaluate the chondrogenic potential of human dermal fibroblasts (DF2 lineage) in two induction protein media: commercial StemPro Differentiation Kit and laboratory-developed media containing TGF-β3 recombinant protein. Methods: The assessment of chondrogenic potential was based on extracellular matrix synthesis. The expression of collagen type I and type II genes (Col1a1, Col2a1) was measured using the RT-qPCR technique. The level of glycosaminoglycan (cartilage matrix proteoglycan) production was assessed at three time points using Alcian blue histochemical staining, followed by statistical evaluation in ImageJ software. Results and Discussion: An increase in the production of extracellular matrix, which is characteristic of hyaline cartilage, in DF2 cell line after induction of chondrogenic differentiation, was confirmed. Conclusions: Consequently, DF2 is considered an applicable cell line for hyaline cartilage tissue engineering due to relatively non-invasive methods of obtaining cell culture, undemanding cultivating conditions, and the possibility of its use in allogeneic grafts.

AB - Abstract: Background: Existing methods of repairing articular cartilage used in medical practice are not fully effective, and therefore, the development of new approaches is a high priority for regenerative medicine. One promising approach is the use of tissue engineering techniques, which involve the use of biomedical cellular products containing biodegradable scaffolds and modified cells. Objective: This study aims to evaluate the chondrogenic potential of human dermal fibroblasts (DF2 lineage) in two induction protein media: commercial StemPro Differentiation Kit and laboratory-developed media containing TGF-β3 recombinant protein. Methods: The assessment of chondrogenic potential was based on extracellular matrix synthesis. The expression of collagen type I and type II genes (Col1a1, Col2a1) was measured using the RT-qPCR technique. The level of glycosaminoglycan (cartilage matrix proteoglycan) production was assessed at three time points using Alcian blue histochemical staining, followed by statistical evaluation in ImageJ software. Results and Discussion: An increase in the production of extracellular matrix, which is characteristic of hyaline cartilage, in DF2 cell line after induction of chondrogenic differentiation, was confirmed. Conclusions: Consequently, DF2 is considered an applicable cell line for hyaline cartilage tissue engineering due to relatively non-invasive methods of obtaining cell culture, undemanding cultivating conditions, and the possibility of its use in allogeneic grafts.

KW - DF2

KW - MMSCs

KW - TGF-β3

KW - chondrogenic differentiation

KW - human dermal fibroblasts

KW - multipotent mesenchymal stromal cells

KW - tissue engineering

UR - https://www.mendeley.com/catalogue/e401e7d2-d908-3793-8983-893388a5d651/

U2 - 10.1134/s1990519x25600292

DO - 10.1134/s1990519x25600292

M3 - Article

VL - 19

SP - S49-S58

JO - Cell and Tissue Biology

JF - Cell and Tissue Biology

SN - 1990-519X

IS - Suppl 1

ER -