Chondrogenic commitment of human umbilical cord blood and umbilical cord-derived mesenchymal stem cells induced by the supernatant of chondrocytes: A comparison study

Abstract

Background: Native cartilage has low capacity for regeneration because it has very few progenitor cells. Human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs) and human umbilical cord-derived MSCs (hUC-MSCs) have been employed as promising sources of stem cells for cartilage injury repair. Reproduction of hyaline cartilage from MSCs remains a challenging endeavor. The paracrine factors secreted by chondrocytes possess the capability to induce chondrogenesis from MSCs.

Methods: The conditioned medium derived from chondrocytes was utilized to induce chondrogenic differentiation of hUCB-MSCs and hUC-MSCs. The expression levels of collagen type I alpha 1 chain (Col1a1), collagen type II alpha 1 chain (Col2a1), and SRY-box transcription factor 9 (SOX9) were assessed through quantitative real-time polymerase chain reaction (qRT-PCR), Western blot (WB), and immunofluorescence (IF) assays. To elucidate the mechanism of differentiation, the concentration of transforming growth factor-β1 (TGF-β1) in the conditioned medium of chondrocytes was quantified using enzyme-linked immunosorbent assay (ELISA). Meanwhile, the viability of cells was assessed using Cell Counting Kit-8 (CCK-8) assays.

Results: The expression levels of Col2a1 and SOX9 were found to be higher in induced hUC-MSCs compared to those in induced hUCB-MSCs. The conditioned medium of chondrocytes contained TGF-β1. The CCK-8 assays revealed that the proliferation rate of hUC-MSCs was significantly higher compared to that of hUCB-MSCs.

Conclusions: The chondrogenic potential and proliferation capacity of hUC-MSCs surpass those of hUCB-MSCs, thereby establishing hUC-MSCs as a superior source of seed cells for cartilage tissue engineering.

Keywords

cartilage diseases / chondrogenic differentiation / mesenchymal stem cells

References

[1] Liu Y, Dzidotor G, Le TT, et al. Exercise-induced piezoelectric stimulation for cartilage regeneration in rabbits. Sci Transl Med. 2022;14(627):eabi7282.

[2] Rua LA, Mumme M, Manferdini C, et al. Engineered nasal cartilage for the repair of osteoarthritic knee cartilage defects. Sci Transl Med. 2021;13(609):eaaz4499.

[3] Ding S-L, Zhao X-Y, Xiong W, et al. Cartilage lacuna-inspired microcarriers drive hyaline neocartilage regeneration. Adv Mater. 2023;35(30):e2212114.

[4] Yan X, Yang B, Chen Y, et al. Anti-friction MSCs delivery system improves the therapy for severe osteoarthritis. Adv Mater. 2021;33(52):e2104758.

[5] Li M, Yin H, Yan Z, et al. The immune microenvironment in cartilage injury and repair. Acta Biomater. 2021;140:23-42.

[6] Kim YG, Choi J, Kim K. Mesenchymal stem cell-derived exosomes for effective cartilage tissue repair and treatment of osteoarthritis. Biotechnol J. 2020;15(12):e2000082.

[7] Iaquinta MR, Lanzillotti C, Mazziotta C, et al. The role of microRNAs in the osteogenic and chondrogenic differentiation of mesenchymal stem cells and bone pathologies. Theranostics. 2021;11(13):6573-6591.

[8] Brose TZ, Kubosch EJ, Schmal H, Stoddart MJ, Armiento AR. Crosstalk between mesenchymal stromal cells and chondrocytes: the hidden therapeutic potential for cartilage regeneration. Stem Cell Rev Rep. 2021;17(5):1647-1665.

[9] Chu D-T, Phuong TNT, Le Bao Tien N, et al. An update on the Progress of isolation, culture, storage, and clinical application of human bone marrow mesenchymal stem/stromal cells. Int J Mol Sci. 2020;21(3):708.

[10] Krawczenko A, Klimczak A. Adipose tissue-derived mesenchymal stem/stromal cells and their contribution to angiogenic processes in tissue regeneration. Int J Mol Sci. 2022;23(5):2425.

[11] Furuoka H, Endo K, Sekiya I. Mesenchymal stem cells in synovial fluid increase in number in response to synovitis and display more tissue-reparative phenotypes in osteoarthritis. Stem Cell Res Ther. 2023;14(1):244.

[12] Han YH, Mao YY, Lee KH, et al. Peroxiredoxin II regulates exosome secretion from dermal mesenchymal stem cells through the ISGylation signaling pathway. Cell Commun Signal. 2023;21(1):296.

[13] Huang J, Huang Z, Liang Y, et al. 3D printed gelatin/hydroxyapatite scaffolds for stem cell chondrogenic differentiation and articular cartilage repair. Biomater Sci. 2021;9(7):2620-2630.

[14] Yong-Beom P, Chul-Won H, Jin-A K, et al. Comparison of undifferentiated versus chondrogenic Predifferentiated mesenchymal stem cells derived from human umbilical cord blood for cartilage repair in a rat model. Am J Sports Med. 2019;47(2):451-461.

[15] Tang S, Chen P, Zhang H, et al. Comparison of curative effect of human umbilical cord-derived mesenchymal stem cells and their small extracellular vesicles in treating osteoarthritis. Int J Nanomed. 2021;16:8185-8202.

[16] Nguyen LT, Tran NT, Than UTT, et al. Optimization of culture conditions for the expansion of umbilical cord-derived mesenchymal stem or stromal cell-like cells using Xeno-free culture conditions. Tissue Eng Part C Methods. 2011;17(4):485-493.

[17] Jung S-H, You J-E, Choi S-W, et al. Polycystin-1 enhances Stemmness potential of umbilical cord blood-derived mesenchymal stem cells. Int J Mol Sci. 2021;22(9):4868.

[18] Chen P, Tang S, Li M, et al. Single-cell and spatial transcriptomics decodes Wharton’s jelly-derived mesenchymal stem cells heterogeneity and a subpopulation with wound repair signatures. Adv Sci. 2023;10(4):e2204786.

[19] Yang HY, Song EK, Kang SJ, Kwak WK, Kang JK, Seon JK. Allogenic umbilical cord blood-derived mesenchymal stromal cell implantation was superior to bone marrow aspirate concentrate augmentation for cartilage regeneration despite similar clinical outcomes. Knee Surg Sports Traumatol Arthrosc. 2022;30(1):208-218.

[20] Bertoni L, Jacquet-Guibon S, Branly T, et al. Evaluation of allogeneic bone-marrow-derived and umbilical cord blood-derived mesenchymal stem cells to prevent the development of osteoarthritis in an equine model. Int J Mol Sci. 2021;22(5):2499.

[21] Xi Y, Yue G, Gao S, Ju R, Wang Y. Human umbilical cord blood mononuclear cells transplantation for perinatal brain injury. Stem Cell Res Ther. 2022;13(1):458.

[22] Hernandez JJ, Beaty DE, Fruhwirth LL, Lopes Chaves AP, Riordan NH. Dodging COVID-19 infection: low expression and localization of ACE2 and TMPRSS2 in multiple donor-derived lines of human umbilical cord-derived mesenchymal stem cells. J Transl Med. 2021;19(1):149.

[23] Kim HJ, Cho KR, Jang H, et al. Intracerebroventricular injection of human umbilical cord blood mesenchymal stem cells in patients with Alzheimer’s disease dementia: a phase I clinical trial. Alzheimers Res Ther. 2021;13(1):154.

[24] Stegen S, Rinaldi G, Loopmans S, et al. Glutamine metabolism controls chondrocyte identity and function. Dev Cell. 2020;53(5):530-544.e8.

[25] Kangari P, Talaei-Khozani T, Razeghian-Jahromi I, Razmkhah M. Mesenchymal stem cells: amazing remedies for bone and cartilage defects. Stem Cell Res Ther. 2020;11(1):492.

[26] Kulsirirat T, Honsawek S, Takeda-Morishita M, et al. The effects of andrographolide on the enhancement of chondrogenesis and osteogenesis in human suprapatellar fat pad derived mesenchymal stem cells. Molecules. 2021;26(7):1831.

[27] Jafri MA, Jafri MA, Kalamegam G, et al. Deciphering the Association of Cytokines, chemokines, and growth factors in chondrogenic differentiation of human bone marrow mesenchymal stem cells using an ex vivo osteochondral culture system. Front cell. Dev Biol. 2020;7:380.

[28] Mazor M, Lespessailles E, Best TM, Ali M, Toumi H. Gene expression and chondrogenic potential of cartilage cells: osteoarthritis grade differences. Int J Mol Sci. 2022;23(18):10610.

[29] Zhou X, Wangyang X, Wang Y, et al. LncRNA DNM3OS regulates GREM2 via miR-127-5p to suppress early chondrogenic differentiation of rat mesenchymal stem cells under hypoxic conditions. Cell Mol Biol Lett. 2021;26(1):22.

[30] Peng J, Mao Z, Liu Y, et al. 12-epi-napelline regulated TGF-β/BMP signaling pathway mediated by BMSCs paracrine acceleration against osteoarthritis. Int Immunopharmacol. 2022;113(Part A):109307.

[31] Wang L, Li S, Xiao H, et al. TGF-β1 derived from macrophages contributes to load-induced tendon-bone healing in the murine rotator cuff repair model by promoting chondrogenesis. Bone Joint Res. 2023;12(3):219-230.

[32] Yang Z, Cai Z, Yang C, Luo Z, Bao X. ALKBH5 regulates STAT3 activity to affect the proliferation and tumorigenicity of osteosarcoma via an m6A-YTHDF2-dependent manner. EBioMedicine. 2022;80:104019.

[33] Burger D, Feucht M, Muench LN, Forkel P, Imhoff AB, Mehl J. Good clinical outcomes after patellar cartilage repair with no evidence for inferior results in complex cases with the need for additional patellofemoral realignment procedures: a systematic review. Knee Surg Sports Traumatol Arthrosc. 2022;30(5):1752-1768.

[34] Doyle EC, Wragg NM, Wilson SL. Intraarticular injection of bone marrow-derived mesenchymal stem cells enhances regeneration in knee osteoarthritis. Knee Surg Sports Traumatol Arthrosc. 2020;28(12):3827-3842.

[35] Bunnell BA. Adipose tissue-derived mesenchymal stem cells. World J Stem Cells. 2021;10(12):3433.

[36] Xie M, Zhang Y, Xiong Z, et al. Generation of hyaline-like cartilage tissue from human mesenchymal stromal cells within the self-generated extracellular matrix. Acta Biomater. 2022;149:150-166.

[37] Zhang D, Ying S, Sun P, et al. A TGF-loading hydrogel scaffold capable of promoting chondrogenic differentiation for repairing rabbit nasal septum cartilage defect. Front Bioeng Biotechnol. 2022;10:1057904.

[38] Liu X, Sun HY, Yan D, et al. In vivo ectopic chondrogenesis of BMSCs directed by mature chondrocytes. Biomaterials. 2010;31(36):9406-9414.

[39] Hua Q, Zhang Y, Li H, et al. Human umbilical cord blood-derived MSCs trans-differentiate into endometrial cells and regulate Th17/Treg balance through NF-κB signaling in rabbit intrauterine adhesions endometrium. Stem Cell Res Ther. 2022;13(1):301.

[40] Wang X, Ouyang L, Chen W, Cao Y, Zhang L. Efficient expansion and delayed senescence of hUC-MSCs by microcarrier-bioreactor system. Stem Cell Res Ther. 2023;14(1):284.

[41] Li X, Duan L, Liang Y, Zhu W, Xiong J, Wang D. Human umbilical cord blood-derived mesenchymal stem cells contribute to chondrogenesis in coculture with chondrocytes. Biomed Res Int. 2016;2016:1-9.

[42] Li X, Liang Y, Xiao X, et al. Cell-to-cell culture inhibits dedifferentiation of chondrocytes and induces differentiation of human umbilical cord-derived mesenchymal stem cells. Biomed Res Int. 2019;2019:5871698.

[43] De Kinderen P, Meester J, Loeys B, et al. Differentiation of induced pluripotent stem cells into chondrocytes: Methods and applications for disease modeling and drug discovery. J Bone Miner Res. 2022;37(3):397-410.

[44] Evans CH, Ghivizzani SC, Robbins PD. Osteoarthritis gene therapy in 2022. Cur Opin. Rheumatol. 2023;35(1):37-43.

[45] Kim P, Park J, Lee D-J, et al. Mast4 determines the cell fate of MSCs for bone and cartilage development. Nat Commun. 2022;13(1):3960.

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2024 The Author(s). Animal Models and Experimental Medicine published by John Wiley & Sons Australia, Ltd on behalf of The Chinese Association for Laboratory Animal Sciences.