State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, China.
State Key Laboratory of Stem Cell and Reproductive Biology, National Stem Cell Resource Center, Chinese Academy of Sciences, Beijing, China.
Cell Prolif. 2023 May;56(5):e13466. doi: 10.1111/cpr.13466. Epub 2023 May 17.
Bone defects (BDs), a prevalent clinically refractory orthopaedic disease, presently have no effective treatments. Mesenchymal stem cells (MSCs) can differentiate into osteoblasts and serve as potential seed cells for bone tissue engineering for BD treatment. However, the feasibility of using MSCs as seed cells for bone tissue engineering remains unclear. As a result, the critical issue of large-scale cell-scaffold preparation remains unresolved. In this study, we demonstrated for the first time that human embryonic stem cell-derived MSCs, also known as immunity-and-matrix-regulatory cells (IMRCs), could be inoculated into microcarriers to create osteogenic micro-tissues appropriate for scalable production in 250 mL bioreactor. IMRCs were generally smaller than umbilical cord-derived MSCs (UCMSCs) and could attach, migrate, proliferate and differentiate within the porous microcarriers, whereas UCMSCs could only attach to the surface of microcarriers. Osteogenic micro-tissues generated from IMRCs-seeded microcarriers significantly increased osteocalcin levels after 21 days of differentiation in a bioreactor. Furthermore, the expression levels of osteogenic biomarker genes/proteins such as alkaline phosphatase (ALP), osteocalcin (OCN), runt-related transcription factor 2 (RUNX2), osteopontin (OPN) and osterix (OSX) were significantly higher than osteogenic micro-tissues derived from UCMSCs-seeded microcarriers. Our findings imply that IMRCs could potentially serve as seed cells for the scalable production of osteogenic micro-tissues for BD treatment.
骨缺损（BDs）是一种常见的临床难治性骨科疾病，目前尚无有效治疗方法。间充质干细胞（MSCs）可分化为成骨细胞，可作为骨组织工程治疗 BD 的潜在种子细胞。然而，将 MSCs 用作骨组织工程种子细胞的可行性尚不清楚。因此，大规模细胞-支架制备的关键问题仍未解决。在这项研究中，我们首次证明，人胚胎干细胞衍生的间充质干细胞，也称为免疫和基质调节细胞（IMRCs），可以接种到微载体中，以创建适合在 250 mL 生物反应器中大规模生产的成骨微组织。IMRCs 通常比脐带间充质干细胞（UCMSCs）小，并且可以在多孔微载体中附着、迁移、增殖和分化，而 UCMSCs 只能附着在微载体的表面。在生物反应器中分化 21 天后，由 IMRC 接种的微载体生成的成骨微组织显著增加了骨钙素水平。此外，成骨生物标志物基因/蛋白的表达水平，如碱性磷酸酶（ALP）、骨钙素（OCN）、 runt 相关转录因子 2（RUNX2）、骨桥蛋白（OPN）和骨形成蛋白 2（OSX），明显高于由 UCMSCs 接种的微载体生成的成骨微组织。我们的研究结果表明，IMRCs 可能可作为用于治疗 BD 的成骨微组织的大规模生产的种子细胞。
