Clem Jones Research Centre For Stem Cells & Tissue Regenerative Therapies, Bond University, Gold Coast, QLD, Australia.
J Craniomaxillofac Surg. 2013 Mar;41(2):153-61. doi: 10.1016/j.jcms.2012.07.003. Epub 2012 Aug 9.
The transplantation of human stem cells seeded on biomaterials holds promise for many clinical applications in cranio-maxillo-facial tissue engineering and regenerative medicine. However, stem cell propagation necessary to produce sufficient cell numbers currently utilizes fetal calf serum (FCS) as a growth supplement which may subsequently transmit animal pathogens. Human platelet lysate (HPL) could potentially be utilized to produce clinical-grade stem cell-loaded biomaterials as an appropriate FCS substitute that is in line with clinically-applicable practice. The goal of this study was to investigate whether HPL can be successfully used to propagate human mesenchymal stem cells (HMSCs) seeded on clinically-approved collagen materials under clinically-applicable conditions using FCS as a control. HMSCs were isolated from bone marrow and cultured in the presence of 10% FCS or 10% HPL. Characterization of HMSCs was performed by flow cytometry and through osteogenic and adipogenic differentiation assays. Proliferative capacity of HMSCs on both matrices was investigated by mitochondrial dehydrogenase assays (WST) and tissue coverage scanning electron microscopy (SEM). The isolated HMSC differentiated into osteogenic and adipogenic cells authenticating the multipotentiality of the HMSCs. WST tests and the SEM images demonstrated that HPL was generally superior to FCS in promoting growth of seeded HMSCs. For all other tests HPL supported HMSCs at least equal to FCS. In conclusion, HPL is an effective growth factor to allow expansion of clinical-grade HMSCs on clinically-approved biomaterials for maxillofacial and oral implantology applications.
将种子细胞接种到生物材料上的移植技术,为颅颌面组织工程和再生医学的许多临床应用带来了希望。然而,为了产生足够数量的细胞,目前需要对种子细胞进行增殖,而这一过程通常使用胎牛血清(FCS)作为生长补充剂,这可能会导致动物病原体的传播。人血小板裂解液(HPL)可以作为 FCS 的替代品,用于生产临床级别的载干细胞生物材料,这符合临床应用的实际情况。本研究的目的是研究在临床应用条件下,HPL 是否可以成功地用于增殖接种在临床批准的胶原材料上的人间充质干细胞(HMSC),并以 FCS 作为对照。从骨髓中分离出 HMSC,在 10% FCS 或 10% HPL 的存在下进行培养。通过流式细胞术和成骨及成脂分化实验对 HMSC 进行鉴定。通过线粒体脱氢酶测定法(WST)和组织覆盖扫描电子显微镜(SEM)研究 HMSC 在两种基质上的增殖能力。分离的 HMSC 分化为成骨细胞和成脂细胞,证明了 HMSC 的多能性。WST 试验和 SEM 图像表明,HPL 通常比 FCS 更能促进接种 HMSC 的生长。对于所有其他测试,HPL 对 HMSC 的支持作用至少与 FCS 相当。总之,HPL 是一种有效的生长因子,可以促进临床级 HMSC 在临床批准的生物材料上的扩增,用于颌面和口腔种植学的应用。