Human umbilical cord mesenchymal stem cell transplantation by different pathways and stem cell tracing by bioluminescence imaging in the treatment of ischemic brain injury

doi:10.3969/j.issn.2095-4344.2017.33.025

Abstract

Abstract:

BACKGROUND: Transplantation of human umbilical cord mesenchymal stem cells can effectively promote the recovery of neurological function in animal models of ischemic brain injury. However, it is unclear on the mechanisms underlying the therapeutic actions of human umbilical cord mesenchymal stem cells and the optimal transplantation way. Bioluminescence imaging has been widely used in the field of stem cell transplantation for real-time dynamic monitoring of the proliferation, migration and survival of transplanted stem cells.
OBJECTIVE: To review the studies of human umbilical cord mesenchymal stem cells in the treatment of ischemic brain injury and the recent research progress of bioluminescence imaging in the stem cell transplantation for the treatment of ischemic brain injury.
METHODS: The key words were “human umbilical cord mesenchymal stem cells, brain ischemia, bioluminescence imaging” in English and Chinese, respectively. The first author retrieved PubMed database, Chinese Journal Full-text Database for relevant articles published from January 2011 to December 2016. Literatures with repetitive content and poor relationship were excluded. A total of 98 literatures were initially retrieved, and finally 38 articles met the inclusion criteria.
RESULTS AND CONCLUSION: Human umbilical cord mesenchymal stem cells can be transplanted via the vein, artery, lumbar puncture, stereotactic operation, intraventricular approach in the treatment of ischemic brain injury. They have their own merits and demerits, which mainly focus on stem cell migration range, survival rate, safety and so on. It has been found that bioluminescence imaging technology for the living is characterized by real-time dynamic monitoring, high sensitivity, high-time resolution and easy operation, which can be used to monitor the migration, proliferation and survival of transplanted stem cells in vivo. Therefore, further investigation on the mechanism of human umbilical cord mesenchymal stem cells that are labeled by bioluminescence imaging technology and selection of the optimal transplantation approach is warranted.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Umbilical Cord, Mesenchymal Stem Cells, Brain Ischemia, Tissue Engineering

CLC Number:

R318

Wang Li-xin, Li Hui-ping. Human umbilical cord mesenchymal stem cell transplantation by different pathways and stem cell tracing by bioluminescence imaging in the treatment of ischemic brain injury[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(33): 5407-5412.

Figures/Tables 2

2.1.1 静脉移植近年多项研究将HUCMSCs经尾静脉注入到大脑中动脉缺血再灌注大鼠体内[24，26-27]，结果发现大鼠神经功能得到改善，脑水肿、梗死体积显著下降，通过调控免疫炎性反应发挥神经保护作用，包括降低促炎性因子，升高抗炎性因子，上调免疫炎性反应初期Foxp3 mRNA表达。Li等[5]证实经尾静脉移植HUCMSCs可改善脑缺血小鼠神经功能，缩小梗死区，促进内源性神经再生，并抑制星形胶质细胞活化。张培培等[25]结果表明人脐带间充质干细胞经尾静脉途径移植可以在宿主脑内存活，通过生成神经元样细胞和神经胶质样细胞、促进新生血管形成并分泌神经营养因子等因素促进大鼠神经功能恢复，且未发现明显免疫排斥反应及异常分化细胞，具有较高的安全性。 Guan等[28]选择将HUCMSCs经股静脉注入模型体内，其能够改善脑缺血小鼠神经功能，神经代谢产物变化较大，N-乙酰天冬氨酸(NAA)/磷酸肌酸(Cr)和胆固醇(Cho)/磷酸肌酸(Cr)比值明显升高，乳酸(Lac)/磷酸肌酸(Cr)比值明显降低，提示股静脉移植HUCMSCs可能通过影响缺血区的神经代谢产物而减轻缺血性脑损伤。Zhu等[29]将5×106个HUCMSCs经股静脉注入脑缺血再灌注兔模型体内，发现移植HUCMSCs抑制了早期脑缺血诱导的血清白细胞介素1β，白细胞介素6和肿瘤坏死因子α水平的增加，并显著降低缺血区周围凋亡细胞的百分比，说明HUCMSCs可能通过抑制脑缺血诱导的早期炎症反应及神经细胞凋亡来发挥作用。以上实验研究证实不论是经尾静脉还是股静脉对于HUCMSCs移植治疗缺血性脑损伤都是安全的，可以通过调控免疫炎性反应、重塑神经环路、内源性血管再生、调控神经代谢产物、表达一些内源性营养因子、抑制脑缺血诱导的神经细胞凋亡等多个途径来发挥作用。 2.1.2 颈动脉移植 Karlupia等[30]经颈动脉将脐血单个核细胞、脐血间充质干细胞、脐带基质间充质干细胞分别注入至大脑中动脉闭塞大鼠体内，移植后2周，与假手术组相比，所有3种细胞组和PBS组的缺血半球均观察到脑源性神经营养因子mRNA的表达显著降低，脐血单个核细胞组降低最少。脐血间充质干细胞组观察到白细胞介素2 mRNA和白细胞介素6 mRNA的表达明显降低。该研究证实经颈动脉移植HUCMSCs治疗缺血性脑损伤的安全性和有效性，其在基因水平调控炎症反应从而发挥作用。 2.1.3 经脑立体定向移植国内刘慧纯等[21]将HUCMSCs立体定向移植至大脑中动脉闭塞大鼠损伤侧纹状体，结果显示，HUCMSCs移植后第7天，移植组mNSS评分显著低于PBS组和模型组，提示经脑立体定向移植HUCMSCs早期即可促进损伤脑组织修复，实验中还发现PBS组在损伤后第14天(即移植操作后第7天)mNSS评分较前略有下降，仍高于对照组，说明移植操作可以加重脑组织损伤，而这种损伤随着时间推移可恢复。通过Brdu标记HUCMSCs，发现移植组大鼠损伤中心及周围均可见Brdu染色阳性细胞及Nestin、微管结合蛋白 2、胶原纤维酸性蛋白、Ⅷ因子、血管内皮生长因子免疫组织化学双染阳性细胞，推测HUCMSCs发挥作用的机制可能与分泌神经营养因子、内源性血管再生有关。其他也有研究发现其机制与神经发生、内源性血管再生、调控免疫炎性反应相关[31-33]。 2.1.4 侧脑室移植 Kim等[34]将1×105个HUCMSCs经侧脑室移植至大脑中动脉闭塞幼鼠损伤侧，通过MRI评价梗死病变体积，圆筒实验和旋转实验评价大鼠神经行为学，结果发现，移植组脑缺血体积缩小，神经功能改善。在缺血半暗带中观察到脱氧核糖核苷酸末端转移酶(TUNEL)、反应性小胶质细胞(ED-1)和胶质纤维酸性蛋白(GFAP)标记的细胞显著增加，这表明HUCMSCs可经神经血管发生及神经保护途径来发挥作用。 2.1.5 不同移植途径比较对HUCMSCs经不同移植途径治疗缺血性脑损伤进行比较，如Lim等[23]分别将1×106、5×105两个剂量HUCMSCs经尾静脉、腰椎穿刺分别注入大脑中动脉闭塞大鼠缺血损伤侧，通过TTC染色和MRI评价梗死病变体积，旋转试验和移除黏附物能力测试评价大鼠神经行为学，结果表明两种移植方式注射1×106个HUCMSCs均可有效改善神经功能，5×105个HUCMSCs鞘内给药显著减少了缺血性损伤，但在静脉注射治疗组中没有显著效果。而经腰椎穿刺移植的HUCMSCs存活时间更长，迁移范围更广泛，并可分化为神经细胞和星形胶质细胞。现有的多项研究支持HUCMSCs经不同移植途径治疗缺血性脑损伤的安全性及有效性。经脑立体定向移植操作可以加重脑损伤，是由于其可能影响梗死灶周围的血供或直接损害脑组织还是其他原因并未阐明；经静脉及腰椎途径移植对比可发现后者存活时间及迁移范围皆优于前者，可能是因为移植的HUCMSCs可以随脑脊液循环迁移到整个大脑和脊髓。目前尚不能明确HUCMSCs治疗缺血性脑损伤的机制，且HUCMSCs各种移植途径之间缺少对比，因此，迫切需要能够活体内动态监测移植干细胞增殖、分化、迁移及作用机制的成像技术。 2.2 活体生物发光成像技术在干细胞治疗缺血性脑损伤中的应用目前BLI可用于监测干细胞移植后的迁移、分布、生存时间，追踪不同因子如营养因子及免疫炎性因子的动态变化，以了解干细胞发挥作用的机制等(图1)，在干细胞治疗缺血性脑损伤领域中广泛应用。"

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