Brzóska Edyta, Grabowska Iwona, Hoser Grazyna, Stremińska Władysława, Wasilewska Danuta, Machaj Eugeniusz Krzysztof, Pojda Zygmunt, Moraczewski Jerzy, Kawiak Jerzy
Department of Clinical Cytology, Medical Center of Postgraduate Education, Warsaw, Poland.
Exp Hematol. 2006 Sep;34(9):1262-70. doi: 10.1016/j.exphem.2006.05.009.
In this report, we demonstrate the participation of human cord blood (HUCB) stem cells in the skeletal muscle regeneration of SCID (severe combined immunodeficient) mice.
The HUCB cells were labeled with the PKH26 fluorescent marker or recognized by an anti-HLA-ABC or anti-beta-2-microglobulin antibody. The HUCB cells were implanted directly into the damaged mouse muscle. The regeneration process and the implanted HUCB cells were traced each day after the damage, throughout a period of 7 days, and additionally at day 30 with the use of flow cytometry and confocal microscopy.
The PKH26-labeled cells isolated from the regenerating muscle were positive for the anti-HLA-ABC antibody. The percentage of the PKH26(+) and HLA-ABC(+) cells decreased from day 1 to day 5. In the regenerating muscle, the percentage of the HLA-ABC(+) cells increased, as measured on days 7 and 30. Moreover, myofibers containing fragments of the PKH26-labeled sarcolemma were noticed. Labeling with the anti-human beta(2)-microglobulin antibody showed the presence of positive cells and myofibers at day 7 of the regeneration, suggesting fusion of human and mouse cells.
We suggest that the HUCB cells implanted into the damaged muscle are present there for at least 30 days and that they participate in the muscle regeneration. Moreover, our study shows that the implanted HUCB cells form human muscle precursor cells residing in the repaired mouse muscle. We suggest that the HUCB cell circulation after transplantation depends on SDF-1 (stromal-derived factor-1) expression in regenerating muscle.
在本报告中,我们展示了人脐带血(HUCB)干细胞参与重症联合免疫缺陷(SCID)小鼠的骨骼肌再生。
用PKH26荧光标记物标记HUCB细胞,或用抗HLA-ABC或抗β2-微球蛋白抗体识别。将HUCB细胞直接植入受损的小鼠肌肉中。损伤后每天追踪再生过程和植入的HUCB细胞,持续7天,并在第30天另外使用流式细胞术和共聚焦显微镜进行追踪。
从再生肌肉中分离出的PKH26标记细胞对抗HLA-ABC抗体呈阳性。PKH26(+)和HLA-ABC(+)细胞的百分比从第1天到第5天下降。在再生肌肉中,在第7天和第30天测量时,HLA-ABC(+)细胞的百分比增加。此外,还发现了含有PKH26标记肌膜片段的肌纤维。用抗人β(2)-微球蛋白抗体标记显示,在再生的第7天存在阳性细胞和肌纤维,提示人和小鼠细胞融合。
我们认为植入受损肌肉中的HUCB细胞在那里至少存在30天,并参与肌肉再生。此外,我们的研究表明,植入的HUCB细胞形成了存在于修复后的小鼠肌肉中的人肌肉前体细胞。我们认为移植后HUCB细胞的循环取决于再生肌肉中基质衍生因子-1(SDF-1)的表达。