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首页> 外文期刊>BioResearch open access. >Bone Progenitors Produced by Direct Osteogenic Differentiation of the Unprocessed Bone Marrow Demonstrate High Osteogenic Potential In Vitro and In Vivo
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Bone Progenitors Produced by Direct Osteogenic Differentiation of the Unprocessed Bone Marrow Demonstrate High Osteogenic Potential In Vitro and In Vivo

机译：未经处理的骨髓直接成骨分化产生的骨祖细胞显示体内和体外的高成骨潜能。

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Tissue-engineered bone grafts seeded with mesenchymal stem cells (MSCs) have been sought as a replacement for bone grafts currently used for bone repair. For production of osteogenic constructs, MSCs are isolated from bone marrow (BM) or other tissues, expanded in culture, then trypsinized, and seeded on a scaffold. Predifferentiation of seeded cells is often desired. We describe here bone progenitor cells (BPCs) obtained by direct osteogenic differentiation of unprocessed BM bypassing isolation of MSCs. Human BM aspirates were incubated for 2 weeks with a commonly used osteogenic medium (OM), except no fetal calf serum, serum substitutes, or growth factors were added, because responding stem and/or progenitor cells were present in the BM milieu. The adherent cells remaining after the culture medium and residual BM were washed out, expressed high levels of bone-specific alkaline phosphatase (ALP) on their surface, demonstrated high ALP activity, were capable of mineralization of the intercellular space, and expressed genes associated with osteogenesis. These parameters in BPCs were similar and even at higher levels compared to MSCs subjected to osteogenic differentiation for 2 weeks. The yield of BPCs per 1?mL BM was 0.71±0.39×106. In comparison, the yield of MSCs produced by adhesion of mononuclear cells derived from the same amount of BM and cultured in a commercial growth medium for 2 weeks was 0.3±0.17×106. When a scaffold was added to the BM-OM mixture, and the mixture was cultured in a simple rotational bioreactor; the resulting BPCs were obtained already seeded on the scaffold. BPCs seeded on scaffolds were capable of proliferation for at least 6 weeks, keeping high levels of ALP activity, expressing osteogenic genes, and mineralizing the scaffolds. Autologous rat BPCs seeded on various scaffolds were transplanted into critical-size calvarial defects. Six weeks after transplantation of polylactic acid/polyglycolic acid scaffolds, 76.1%±18.3% of the defects were filled with a new bone, compared to 37.9%±28.4% in the contralateral defects transplanted with the scaffolds without cells.
机译：已经寻求了植入间充质干细胞（MSC）的组织工程化骨移植物，以替代目前用于骨修复的骨移植物。为了生产成骨构建体，从骨髓（BM）或其他组织中分离MSC，在培养物中扩增，然后用胰蛋白酶消化，然后接种在支架上。通常需要种子细胞的预分化。我们在这里描述了未经处理的BM绕过MSC分离的直接成骨分化获得的骨祖细胞（BPC）。将人BM抽吸物与常用的成骨培养基（OM）孵育2周，但不添加胎牛血清，血清替代物或生长因子，因为BM环境中存在有反应的干细胞和/或祖细胞。洗去培养基和残留的BM后残留的贴壁细胞，在其表面表达高水平的骨特异性碱性磷酸酶（ALP），表现出高的ALP活性，能够矿化细胞间空间，并表达与成骨作用。与经过2周成骨分化的MSC相比，BPC中的这些参数相似，甚至更高。每1？mL BM的BPC产量为0.71±0.39×10 6 。相比之下，由等量BM衍生并在商业生长培养基中培养2周的单核细胞粘附产生的MSC的产量为0.3±0.17×10 6 。当向BM-OM混合物中添加支架时，将混合物在简单的旋转生物反应器中培养;得到的BPC已经被接种在支架上。接种在支架上的BPC能够增殖至少6周，保持高水平的ALP活性，表达成骨基因，并使支架矿化。种植在各种支架上的自体大鼠BPC被移植到临界大小的颅骨缺损中。聚乳酸/聚乙醇酸支架移植六周后，新骨填充了76.1％±18.3％的缺损，而无细胞支架移植的对侧缺损为37.9％±28.4％。

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