Human umbilical cord blood-derived mesenchymal stem cells in the cultured rabbit intervertebral disc: a novel cell source for disc repair.
Human umbilical cord blood-derived mesenchymal stem cells in the cultured rabbit intervertebral disc: a novel cell source for disc repair.
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培养的兔椎间盘中的人脐带血液衍生的间充质干细胞：椎间盘修复的新细胞来源。
DOI：
10.1097/phm.0b013e31825f148a
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发表时间：
2013-05
影响因子：
3
通讯作者：
Zhang Y
中科院分区：
文献类型：
--
作者：
Anderson DG;Markova D;An HS;Chee A;Enomoto-Iwamoto M;Markov V;Saitta B;Shi P;Gupta C;Zhang Y
与症状性椎间盘退变相关的背痛是一种常见的临床病症。椎间盘（IVD）细胞凋亡和衰老随着衰老和退化而增加。用能够产生和维持细胞外基质的细胞重新填充 IVD 将是手术的替代疗法。本研究的目的是确定人脐带血来源的间充质干细胞 (hUCB-MSC) 作为椎间盘修复的新型细胞来源的潜力。在本研究中，我们旨在确认 hUCB-MSC 在微团培养和注射到兔 IVD 外植体培养物中后分化和显示软骨细胞样表型的潜力。我们还想确认 hUCB-MSC 移植到 IVD 外植体培养物中后的存活率。这项研究包括微团培养和体外兔 IVD 外植体培养，以评估 hUCB-MSC 存活和分化以显示软骨细胞样表型。首先，在微团中培养 hUCB-MSC，并用阿尔新蓝染料染色。其次，为了确认细胞存活，在注射到整个兔 IVD 外植体（宿主）之前，用红外染料和荧光染料标记 hUCB-MSC。然后将 IVD 外植体培养 4 周。通过两种独立的技术确认细胞存活：在器官水平检测红外染料的成像系统和在细胞水平检测荧光染料的荧光显微镜。通过用 CellTracker green（一种针对活细胞的膜渗透示踪剂）对外植体进行染色来评估细胞活力。通过聚合酶链反应评估人类 II 型胶原蛋白基因表达（来自移植物）。我们已经证明，在微团中培养的 hUCB-MSC 被阿尔新蓝染料染成蓝色，这表明产生了富含蛋白聚糖的细胞外基质。在培养的兔 IVD 外植体中，hUCB-MSC 存活至少 4 周并表达人 II 型胶原基因，表明注射的 hUCB-MSC 正在分化为软骨细胞样谱系。这项研究证明了 hUBC-MSC 在注射到兔 IVD 中时能够存活并呈现软骨细胞样表型。这些数据支持 hUBC-MSC 作为椎间盘修复细胞来源的潜力。在人体试验之前，需要进一步测量宿主对注射的反应并在动物模型中进行研究。
Back pain associated with symptomatic disc degeneration is a common clinical condition. Intervertebral disc (IVD) cell apoptosis and senescence increase with aging and degeneration. Repopulating the IVD with cells that could produce and maintain extracellular matrix would be an alternative therapy to surgery. The objective of this study was to determine the potential of human umbilical cord blood–derived mesenchymal stem cells (hUCB-MSCs) as a novel cell source for disc repair. In this study, we intended to confirm the potential for hUCB-MSCs to differentiate and display a chondrocyte-like phenotype after culturing in micromass and after injection into the rabbit IVD explant culture. We also wanted to confirm hUCB-MSC survival after transplantation into the IVD explant culture. This study consisted of micromass cultures and in vitro rabbit IVD explant cultures to assess hUCB-MSC survival and differentiation to display chondrocyte-like phenotype. First, hUCB-MSCs were cultured in micromass and stained with Alcian blue dye. Second, to confirm cell survival, hUCB-MSCs were labeled with an infrared dye and a fluorescent dye before injection into whole rabbit IVD explants (host). IVD explants were then cultured for 4 wks. Cell survival was confirmed by two independent techniques: an imaging system detecting the infrared dye at the organ level and fluorescence microscopy detecting fluorescent dye at the cellular level. Cell viability was assessed by staining the explant with CellTracker green, a membrane-permeant tracer specific for live cells. Human type II collagen gene expression (from the graft) was assessed by polymerase chain reaction. We have shown that hUCB-MSCs cultured in micromass are stained blue with Alcian blue dye, which suggests that proteoglycan-rich extracellular matrix is produced. In the cultured rabbit IVD explants, hUCB-MSCs survived for at least 4 wks and expressed the human type II collagen gene, suggesting that the injected hUCB-MSCs are differentiating into a chondrocyte-like lineage. This study demonstrates the abiity of hUBC-MSCs to survive and assume a chondrocyte-like phenotype when injected into the rabbit IVD. These data support the potential for hUBC-MSCs as a cell source for disc repair. Further measures of the host response to the injection and studies in animal models are needed before trials in humans.