The Shraga Segal Department of Microbiology and Immunology, Faculty of Health Sciences, Ben-Gurion University, Beer Sheva, Israel.
Cytotherapy. 2009;11(7):837-48. doi: 10.3109/14653240903136979.
Telomerase is a ribonucleoprotein that maintains the length of telomeres, and thus controls the proliferation and lifespan of cells. Recent studies suggest the involvement of telomerase in the protection of cells from apoptosis. Adult human mesenchymal stromal cells (hMSC) possess the capacity to proliferate and differentiate into a variety of cell types. hMSC derived from a healthy donor lack telomerase activity but their expression has not been investigated in hMSC derived from diseased adults. Cell replacement therapy using adult hMSC has been suggested as a promising therapeutic approach for amyotrophic lateral sclerosis (ALS). Therefore, we characterized the telomerase activity and expression in hMSC derived from bone marrow (BM) of ALS patients and compared them with those derived from a healthy donor.
Telomerase activity was examined with a TRAP assay and real-time polymerase chain reaction telomerase quantification assay. Telomerase protein was detected by Western blot and immunofluorescence analysis, and telomerase RNA transcripts were identified by Northern blot.
Telomerase activity, telomerase enzyme protein and telomerase RNA transcripts were demonstrated in hMSC derived from ALS, but were undetectable in hMSC from the healthy donor. Telomerase activity in the hMSC of ALS patients was 106-fold lower compared with tumor cells.
The detection of telomerase expression in hMSC derived from ALS patients and not a healthy donor suggests a possible role for telomerase in the response of hMSC to the disease. The presence of telomerase expression did not impair the ability of the ALS hMSC to differentiate, suggesting the use of these cells for cytotherapy treatments.
端粒酶是一种核糖核蛋白，可维持端粒的长度，从而控制细胞的增殖和寿命。最近的研究表明，端粒酶参与了细胞免受凋亡的保护。成人间充质基质细胞（hMSC）具有增殖和分化为多种细胞类型的能力。来自健康供体的 hMSC 缺乏端粒酶活性，但尚未研究过来自患病成年人的 hMSC 中的表达。使用成年 hMSC 的细胞替代疗法已被提议作为肌萎缩侧索硬化症（ALS）的有前途的治疗方法。因此，我们对来自 ALS 患者骨髓（BM）的 hMSC 中的端粒酶活性和表达进行了表征，并将其与来自健康供体的 hMSC 进行了比较。
使用 TRAP 分析和实时聚合酶链反应端粒酶定量分析检测端粒酶活性。通过 Western blot 和免疫荧光分析检测端粒酶蛋白，通过 Northern blot 鉴定端粒酶 RNA 转录本。
在来自 ALS 的 hMSC 中检测到端粒酶活性、端粒酶酶蛋白和端粒酶 RNA 转录本，但在来自健康供体的 hMSC 中无法检测到。与肿瘤细胞相比，ALS 患者的 hMSC 中端粒酶活性低 106 倍。
在来自 ALS 患者而非健康供体的 hMSC 中检测到端粒酶表达表明端粒酶在 hMSC 对疾病的反应中可能起作用。端粒酶表达的存在并未损害 ALS hMSC 分化的能力，这表明这些细胞可用于细胞治疗。
