Schultz Sherri S, Abraham Sherly, Lucas Paul A
Department of Pathology, New York Medical College, Valhalla, New York 10595, USA.
Wound Repair Regen. 2006 Mar-Apr;14(2):224-31. doi: 10.1111/j.1743-6109.2006.00114.x.
Adult stem cells capable of differentiating into phenotypes from all three dermal layers were isolated from adult rat muscle. Stem cells were obtained by enzymatic digestion, followed by primary culture in Eagle's minimum essential medium +10% preselected horse serum. When the cells reached confluence, they were released by trypsin, filtered to remove differentiated myotubes, and then slow frozen in 7.5% dimethylsulfoxide to -80 degrees C. Thawed cells were the stem cells and were induced to differentiate with the nonspecific differentiating agent dexamethasone at concentrations of 10(-10)-10(-6) M. After a 6-week treatment with dexamethasone, the cells were assayed by immunohistochemistry for phenotypes of the mesodermal, ectodermal, and endodermal lineages. Examples of mesodermal phenotypes identified were as follows: bone, cartilage, and skeletal, smooth, and cardiac muscle. Ectodermal phenotypes identified were as follows: neurons and oligodendrocytes. Hepatocyte phenotypes identified represented the endodermal lineage. All the phenotypes were observed only with treatment with dexamethasone. However, nestin was observed in the absence of dexamethasone and may be a marker for uncommitted pluripotent stem cells. The results show that adult muscle contains pluripotent stem cells capable of differentiating across all three dermal lineages. Such cells could be used in the context of tissue engineering.
从成年大鼠肌肉中分离出了能够分化为所有三层真皮层表型的成体干细胞。通过酶消化获得干细胞，然后在伊格尔最低必需培养基+10%预先筛选的马血清中进行原代培养。当细胞达到汇合状态时，用胰蛋白酶将其消化，过滤以去除分化的肌管，然后在7.5%二甲基亚砜中缓慢冷冻至-80℃。解冻后的细胞即为干细胞，并用浓度为10(-10)-10(-6)M的非特异性分化剂地塞米松诱导其分化。用地塞米松处理6周后，通过免疫组织化学法检测细胞中中胚层、外胚层和内胚层谱系的表型。鉴定出的中胚层表型示例如下：骨、软骨以及骨骼肌、平滑肌和心肌。鉴定出的外胚层表型如下：神经元和少突胶质细胞。鉴定出的肝细胞表型代表内胚层谱系。所有表型仅在用地塞米松处理时观察到。然而，在没有地塞米松的情况下观察到了巢蛋白，它可能是未定向多能干细胞的标志物。结果表明，成年肌肉中含有能够跨所有三个真皮谱系分化的多能干细胞。这类细胞可用于组织工程领域。
