Pravin J Mishra, Prasun J Mishra, Laboratory of Cancer Biology and Genetics, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892-4264, United States.
World J Stem Cells. 2012 May 26;4(5):35-43. doi: 10.4252/wjsc.v4.i5.35.
To compare the efficacy of cell-free derivatives from Bone marrow derived human mesenchymal stem cells (hMSCs) in wound therapy.
hMSCs have been shown to play an important role in wound therapy. The present study sought to compare efficacy of hMSCs and cell-free derivatives of hMSCs, which may be clinically more relevant as they are easier to prepare, formulate and transport. hMSCs were isolated from human bone marrow and cultured. Multi lineage differentiation of hMSCs was performed to confirm their identity. The ability of hMSCs to migrate was evaluated using in vitro and in vivo migration assays. Cell lysates and conditioned medium concentrate was prepared from hMSCs (see Methods for details). Wounds were induced in mice and wound areas were measure before and after cell and cell-free derivative treatment. RNA and proteins were extracted from the skin and cytokine levels were measured.
Co-culture of hMSCs with keratinocytes resulted in increased expression of CXCL-12 (SDF1) and ENA78 (CXCL-5) in the conditioned media indicating that the hMSCs can respond to signals from keratinocytes. Accelerated wound closure was observed when hMSCs were injected near the site of excisional wounds in athymic as well as NOD/SCID mice. Interestingly, cell-free lysates prepared from hMSCs were also effective in inducing accelerated wound closure and increased expression of SDF1 and CXCL-5 at the wound bed. Additionally, concentrated media from hMSCs as well as an emulsion containing lysates prepared from hMSCs was also found to be more effective in rapid re-epithelialization than fibroblasts or vehicle-alone control. Use of cell-free derivatives may help replace expensive wound care approaches including use of growth factors, epidermal/dermal substitutes, synthetic membranes, cytokines, and matrix components, and most importantly avoid transmission of pathogens from human and animal products.
These results encourage development of derivatives of hMSCs for wound care and re-epithelialization applications.
比较骨髓间充质干细胞（hMSC）无细胞衍生物在伤口治疗中的疗效。
hMSC 在伤口治疗中发挥重要作用。本研究旨在比较 hMSC 和 hMSC 无细胞衍生物的疗效，由于后者更容易制备、配方和运输，因此在临床上可能更相关。从人骨髓中分离出 hMSC 并进行培养。进行多系分化以确认其身份。通过体外和体内迁移实验评估 hMSC 的迁移能力。从 hMSC 中制备细胞裂解物和条件培养基浓缩物（详见方法）。在小鼠中诱导伤口，并在细胞和无细胞衍生物治疗前后测量伤口面积。从皮肤中提取 RNA 和蛋白质，并测量细胞因子水平。
hMSC 与角质形成细胞共培养可导致条件培养基中 CXCL-12（SDF1）和 ENA78（CXCL-5）表达增加，表明 hMSC 可对角质形成细胞的信号做出反应。在无胸腺和 NOD/SCID 小鼠的切创部位附近注射 hMSC 可观察到伤口愈合加速。有趣的是，从 hMSC 制备的无细胞裂解物也可有效诱导伤口快速闭合，并增加伤口床处 SDF1 和 CXCL-5 的表达。此外，与成纤维细胞或单独载体相比，hMSC 浓缩培养基以及含有 hMSC 裂解物的乳剂更有利于快速再上皮化。使用无细胞衍生物可能有助于替代昂贵的伤口护理方法，包括使用生长因子、表皮/真皮替代物、合成膜、细胞因子和基质成分，最重要的是避免从人和动物产品传播病原体。
这些结果鼓励开发 hMSC 的衍生物用于伤口护理和再上皮化应用。
