人类间充质基质细胞的年龄依赖性神经外胚层分化能力：自体细胞替代策略的局限性。

Age-dependent neuroectodermal differentiation capacity of human mesenchymal stromal cells: limitations for autologous cell replacement strategies.

机构信息

Department of Neurology, Dresden University of Technology, Dresden, Germany.

出版信息

Cytotherapy. 2010;12(1):17-30. doi: 10.3109/14653240903313941.

DOI:10.3109/14653240903313941

PMID:19878082

Abstract

BACKGROUND AIMS

Human adult bone marrow (BM)-derived mesenchymal stromal cells (hMSC) are reported to break germ layer commitment and differentiate into cells expressing neuroectodermal properties. Although it is of pivotal interest for cell replacement therapies for neurologic disorders, no data exist on the influence of the donor's age on this multipotent differentiation behavior.

METHODS

We evaluated various epigenetic neuroectodermal conversion protocols in adult hMSC derived from older donors (>45 versus 18-35 years of age) using quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) and immunocytochemistry. The protocols included single- and multi-step conversion-differentiation protocols combined with co-culture techniques. Furthermore, the age dependency of mesodermal differentiation potential and cell senescence were investigated.

RESULTS

The neuroectodermal differentiation potential of hMSC derived from old donors was completely lost, with no cells showing mature neuroectodermal phenotypes using single- and multi-step conversion-differentiation protocols and no improvement of neurogenesis by various co-culture conditions. Comparison of young versus old donor-derived hMSC showed fewer cells expressing early neuroectodermal marker proteins in the latter samples. qRT-PCR showed reduced expression of the proliferation marker KI67 and the neuroectodermal gene NES (nestin) in old donor-derived cells compared with young donor hMSC. Telomere length analysis showed no general cell aging.

CONCLUSIONS

Our data provide evidence that only young donor-derived hMSC can be epigenetically differentiated in vitro into neuroectodermal cells, pointing towards senescence of multipotentiality of old donor-derived hMSC. There is thus an urgent need to develop better protocols for successful neuroectodermal differentiation of hMSC from old individuals as a prerequisite for autologous cell replacement strategies for neurologic diseases in elderly patients.

摘要

背景目的

据报道，人类成人骨髓（BM）来源的间充质基质细胞（hMSC）可打破胚层承诺，并分化为表达神经外胚层特性的细胞。尽管这对于神经紊乱的细胞替代治疗具有重要意义，但关于供体年龄对这种多能分化行为的影响尚无数据。

方法

我们使用定量逆转录聚合酶链反应（qRT-PCR）和免疫细胞化学评估了来自年龄较大的供体（>45 岁与 18-35 岁）的成人 hMSC 的各种表观遗传神经外胚层转化方案。该方案包括单一和多步转化-分化方案，以及共培养技术。此外，还研究了中胚层分化潜能和细胞衰老的年龄依赖性。

结果

来自老年供体的 hMSC 的神经外胚层分化潜能完全丧失，使用单一和多步转化-分化方案均未观察到成熟的神经外胚层表型的细胞，并且各种共培养条件均未改善神经发生。与年轻供体衍生的 hMSC 相比，老年供体衍生的 hMSC 中表达早期神经外胚层标记蛋白的细胞更少。qRT-PCR 显示，与年轻供体 hMSC 相比，老年供体衍生的细胞中增殖标记物 KI67 和神经外胚层基因 NES（巢蛋白）的表达降低。端粒长度分析显示，老年供体衍生的细胞没有普遍的细胞衰老。