背景:沃顿的果冻(WJ)间充质基质细胞(MSC)已成为许多适应症的有吸引力的同种异体疗法,除了骨相关的条件需要新的组织形成。这可以用MSC的明显顽抗来解释,WJ在体外分化为成骨谱系,与允许的骨髓(BM)来源的MSC(MSC,BM)容易进入骨细胞。因此,MSC的实际体内成骨能力,WJ正在讨论中。
方法:我们研究了生理骨环境如何在体外和体内影响顽固性MSCs的成骨作用。为此,将BM和WJ来源的MSC共培养并使用transwell在体外诱导同步成骨分化。对于体内实验,免疫缺陷小鼠注射单剂量人MSC,6周后评估骨形成.
结果:MSC的共同培养,BM和MSC,WJ在三周后导致两种细胞类型的有效骨生成。然而,MSC,在没有MSC的情况下,WJ未能进入骨细胞,BM的成骨刺激。体内研究表明,在用MSC治疗的62.5%的小鼠中,胫骨髓腔内的骨形成成功。WJ.相比之下,仅在25%的MSC中观察到新形成的小梁,BM处理的小鼠。人COXIV的免疫组织化学染色显示注入细胞在注射部位的持久性。此外,人类来源的细胞也在大脑中被发现,心,脾,脾用工程MSC治疗的一些动物的肾脏和性腺,WJ(eMSC,WJ).重要的是,没有宏观组织病理学改变,在MSC治疗的小鼠中检测到异位骨形成或任何其他不良事件.
结论:我们的研究结果表明,在生理骨微环境中,MSC的成骨承诺,WJ与MSC相当,BM,并支持使用现成的同种异体MSC,WJ产品在骨修复和骨再生方面的应用。
Wharton\'s Jelly (WJ) Mesenchymal Stromal Cells (MSC) have emerged as an attractive allogeneic therapy for a number of indications, except for bone-related conditions requiring new tissue formation. This may be explained by the apparent recalcitrance of MSC,WJ to differentiate into the osteogenic lineage in vitro, as opposed to permissive bone marrow (BM)-derived MSCs (MSC,BM) that readily commit to bone cells. Consequently, the actual osteogenic in vivo capacity of MSC,WJ is under discussion.
We investigated how physiological bone environments affect the osteogenic commitment of recalcitrant MSCs in vitro and in vivo. To this end, MSC of BM and WJ origin were co-cultured and induced for synchronous osteogenic differentiation in vitro using transwells. For in vivo experiments, immunodeficient mice were injected intratibially with a single dose of human MSC and bone formation was evaluated after six weeks.
Co-culture of MSC,BM and MSC,WJ resulted in efficient osteogenesis in both cell types after three weeks. However, MSC,WJ failed to commit to bone cells in the absence of MSC,BM\'s osteogenic stimuli. In vivo studies showed successful bone formation within the medullar cavity of tibias in 62.5% of mice treated with MSC, WJ. By contrast, new formed trabeculae were only observed in 25% of MSC,BM-treated mice. Immunohistochemical staining of human COXIV revealed the persistence of the infused cells at the site of injection. Additionally, cells of human origin were also identified in the brain, heart, spleen, kidney and gonads in some animals treated with engineered MSC,WJ (eMSC,WJ). Importantly, no macroscopic histopathological alterations, ectopic bone formation or any other adverse events were detected in MSC-treated mice.
Our findings demonstrate that in physiological bone microenvironment, osteogenic commitment of MSC,WJ is comparable to that of MSC,BM, and support the use of off-the-shelf allogeneic MSC,WJ products in bone repair and bone regeneration applications.