Department of Orthopedic Surgery, School of Medicine, Stanford University, Stanford, California, USA.
Gladstone Institute of Neurological Disease, San Francisco, California, USA.
Tissue Eng Part A. 2022 May;28(9-10):433-446. doi: 10.1089/ten.TEA.2021.0172. Epub 2021 Dec 31.
Mesenchymal stromal cells (MSCs) have been widely investigated for their regenerative capacity, anti-inflammatory properties and beneficial immunomodulatory effects across multiple clinical indications. Nevertheless, their widespread clinical utilization is limited by the variability in MSC quality, impacted by donor age, metabolism, and disease. Human induced pluripotent stem cells (hiPSCs) generated from readily accessible donor tissues, are a promising source of stable and rejuvenated MSC but differentiation methods generally require prolonged culture and result in low frequencies of stable MSCs. To overcome this limitation, we have optimized a quick and efficient method for hiPSC differentiation into footprint-free MSCs (human induced MSCs [hiMSCs]) in this study. This method capitalizes on the synergistic action of growth factors Wnt3a and Activin A with bone morphogenetic protein-4 (BMP4), leading to an enrichment of MSC after only 4 days of treatment. These hiMSCs demonstrate a significant upregulation of mesenchymal stromal markers (CD105, CD90, CD73, and cadherin 11) compared with bone marrow-derived MSCs (bmMSCs), with reduced expression of the pluripotency genes (octamer-binding transcription factor [], cellular myelocytomatosis oncogene []]) compared with hiPSC. Moreover, they show improved proliferation capacity in culture without inducing any teratoma formation . Osteogenesis, chondrogenesis, and adipogenesis assays confirmed the ability of hiMSCs to differentiate into the three different lineages. Secretome analyses showed cytokine profiles compared with bmMSCs. Encapsulated hiMSCs in alginate beads cocultured with osteoarthritic (OA) cartilage explants showed robust immunomodulation, with stimulation of cell growth and proteoglycan production in OA cartilage. Our quick and efficient protocol for derivation of hiMSC from hiPSC, and their encapsulation in microbeads, therefore, presents a reliable and reproducible method to boost the clinical applications of MSCs.
间充质基质细胞（MSCs）因其在多个临床适应症中的再生能力、抗炎特性和有益的免疫调节作用而被广泛研究。然而，由于 MSC 质量的可变性，受供体年龄、代谢和疾病的影响，其广泛的临床应用受到限制。从易于获得的供体组织中产生的人类诱导多能干细胞（hiPSCs）是稳定和恢复活力的 MSC 的有前途的来源，但分化方法通常需要长时间培养，并且导致稳定 MSC 的频率较低。为了克服这一限制，我们在本研究中优化了一种快速有效的 hiPSC 分化为无足迹 MSC（人诱导 MSC [hiMSCs]）的方法。该方法利用生长因子 Wnt3a 和激活素 A 与骨形态发生蛋白-4（BMP4）的协同作用，仅在 4 天的治疗后就导致 MSC 的富集。与骨髓来源的 MSC（bmMSCs）相比，这些 hiMSCs 表现出间充质基质标记物（CD105、CD90、CD73 和钙黏蛋白 11）的显著上调，与 hiPSC 相比，多能性基因（八聚体结合转录因子 []、细胞髓细胞瘤癌基因 []）的表达降低。此外，它们在培养中显示出改善的增殖能力，而不会诱导任何畸胎瘤形成。成骨、软骨和成脂分化试验证实了 hiMSCs 分化为三种不同谱系的能力。分泌组分析显示与 bmMSCs 相比的细胞因子谱。与骨关节炎（OA）软骨外植体共培养的藻酸盐珠包封的 hiMSCs 显示出强大的免疫调节作用，刺激 OA 软骨中的细胞生长和糖蛋白聚糖的产生。因此，我们从 hiPSC 快速有效地获得 hiMSC 的方案及其在微珠中的包封，为促进 MSC 的临床应用提供了一种可靠且可重复的方法。
