Clumps of Mesenchymal Stem Cell/Extracellular Matrix Complexes Generated with Xeno-Free Conditions Facilitate Bone Regeneration via Direct and Indirect Osteogenesis

• Published in: International journal of molecular sciences Vol. 20; no. 16; p. 3970
• Main Authors: Motoike, Souta, Kajiya, Mikihito, Komatsu, Nao, Horikoshi, Susumu, Ogawa, Tomoya, Sone, Hisakatsu, Matsuda, Shinji, Ouhara, Kazuhisa, Iwata, Tomoyuki, Mizuno, Noriyoshi, Fujita, Tsuyoshi, Ikeya, Makoto, Kurihara, Hidemi
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 15.08.2019; MDPI
• Subjects: 3D culture; Animals; Biodegradability; Biomaterials; Biomedical materials; Bone growth; Bone marrow; Bone mass; bone regeneration; Bone Regeneration - genetics; Bone Regeneration - physiology; Cell Differentiation - physiology; Clumps; Defects; Extracellular matrix; Extracellular Matrix - metabolism; Implantation; Inflammation; Male; Mesenchymal stem cells; Mesenchymal Stem Cells - metabolism; Mice; Mice, SCID; MSCs; Osteogenesis; Osteogenesis - physiology; Proteins; Regeneration; Regeneration (physiology); scaffold-free; Staining; Stem cells; Surgical implants; Tissue Engineering; X-Ray Microtomography; xeno-free; Xenografts; bone regeneration; MSCs; 3D culture; scaffold-free; xeno-free
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms20163970

Three-dimensional clumps of mesenchymal stem cell (MSC)/extracellular matrix (ECM) complexes (C-MSCs) consist of cells and self-produced ECM. We demonstrated previously that C-MSCs can be transplanted into bone defect regions with no artificial scaffold to induce bone regeneration. To apply C-MSCs in a clinical setting as a reliable bone regenerative therapy, the present study aimed to generate C-MSCs in xeno-free/serum-free conditions that can exert successful bone regenerative properties and to monitor interactions between grafted cells and host cells during bone healing processes. Human bone marrow-derived MSCs were cultured in xeno-free/serum-free medium. To obtain C-MSCs, confluent cells that had formed on the cellular sheet were scratched using a micropipette tip and then torn off. The sheet was rolled to make a round clump of cells. Then, C-MSCs were transplanted into an immunodeficient mouse calvarial defect model. Transplantation of C-MSCs induced bone regeneration in a time-dependent manner. Immunofluorescence staining showed that both donor human cells and host mice cells contributed to bone reconstruction. Decellularized C-MSCs implantation failed to induce bone regeneration, even though the host mice cells can infiltrate into the defect area. These findings suggested that C-MSCs generated in xeno-free/serum-free conditions can induce bone regeneration via direct and indirect osteogenesis.