Implanted mouse bone marrow-derived cells reconstruct layered smooth muscle structures in injured urinary bladders
Implanted mouse bone marrow-derived cells reconstruct layered smooth muscle structures in injured urinary bladders
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DOI：
10.3727/096368908784153850
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发表时间：
2008-01-01
影响因子：
3.3
通讯作者：
Nishizawa, Osamu
中科院分区：
文献类型：
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作者：
Imamura, Tetsuya;Kinebuchi, Yoshiaki;Nishizawa, Osamu
这项研究是一项初步调查，以确定骨髓来源的细胞，当植入冷冻损伤的膀胱，分化成平滑肌细胞和重建平滑肌层。从雄性ICR小鼠的股骨收获骨髓细胞，并在胶原包被的培养皿中培养7天。培养5天后，用绿色荧光蛋白（GFP）基因转染细胞，用于在受体组织中鉴定。植入前3天，用干冰冷冻的铁棒损伤雌性裸鼠的后膀胱壁。在培养后7天和损伤后3天，将贴壁的、增殖的GFP标记的骨髓来源的细胞（1.0 × 10（5）个细胞）植入损伤区域。对于对照，注射无细胞溶液。植入后14天，通过组织学、基因表达和膀胱测压研究分析实验膀胱。就在植入前，受损区域没有任何平滑肌层。14天后，用GFP抗体检测移植细胞在受体组织中的存活。植入区域具有由再生的平滑肌标记物阳性细胞组成的明显的平滑肌层。植入的GFP标记的细胞分化成形成层的平滑肌细胞。分化的细胞在植入区域内彼此接触，以及宿主的平滑肌细胞。结果，重建的平滑肌层整合到宿主组织中。注射无细胞溶液的对照组小鼠仅发育了少量平滑肌细胞，没有层。膀胱测量研究表明，植入细胞的小鼠产生了与正常小鼠相似的膀胱收缩，而对照小鼠则没有。总之，小鼠骨髓来源的细胞可以重建受损膀胱中的分层平滑肌结构，以修复泌尿功能障碍。
This study is a preliminary investigation to determine if bone marrow-derived cells, when implanted into freeze-injured urinary bladders, differentiate into smooth muscle cells and reconstruct smooth muscle layers. Bone marrow cells were harvested from femurs of male ICR mice and cultured in collagen-coated dishes for 7 days. After 5 days of culture, the cells were transfected with green fluorescent protein (GFP) genes for identification in recipient tissues. Three days prior to implantation, the posterior urinary bladder walls of female nude mice were injured with an iron bar refrigerated by dry ice. Seven days after the culture and 3 days after the injury, adherent, proliferating GFP-labeled bone marrow-derived cells (1.0 x 10(5) cells) were implanted into the injured regions. For controls, a cell-free solution was injected. At 14 days after implantation, the experimental urinary bladders were analvzed by histological, gene expression, and cystometric investigations. Just prior to implantation, the injured regions did not have any smooth muscle layers. After 14 days, the implanted cells surviving in the recipient tissues were detected with GFP antibody. The implanted regions had distinct smooth muscle layers composed of regenerated smooth muscle marker-positive cells. The implanted GFP-labeled cells differentiated into smooth muscle cells that formed into layers. The differentiated cells contacted each other within the implanted region as well as smooth muscle cells of the host. As a result, the reconstructed smooth muscle layers were integrated into the host tissues. Control mice injected with cell-free solution developed only few smooth muscle cells and no layers. Cystometric investigations showed that mice with implanted the cells developed bladder contractions similar to normal mice, whereas control mice did not. In summary, mouse bone marrow-derived cells can reconstruct layered smooth muscle structures in injured bladders to remediate urinary dysfunction.