胎儿心脏间充质干细胞表达胚胎标志物，并表现出向三个胚层的所有细胞分化的能力。
Fetal cardiac mesenchymal stem cells express embryonal markers and exhibit differentiation into cells of all three germ layers.
机构信息
Garikipati Venkata Naga Srikanth, Naresh Kumar Tripathy, Soniya Nityanand, Stem Cell Research Facility, Department of Hematology, Sanjay Gandhi Post-Graduate Institute of Medical Sciences, Lucknow 226014, India.
出版信息
World J Stem Cells. 2013 Jan 26;5(1):26-33. doi: 10.4252/wjsc.v5.i1.26.
AIM
To study the expression of embryonal markers by fetal cardiac mesenchymal stem cells (fC-MSC) and their differentiation into cells of all the germ layers.
METHODS
Ten independent cultures of rat fC-MSC were set up from cells derived from individual or pooled fetal hearts and studies given below were carried out at passages 3, 6, 15 and 21. The phenotypic markers CD29, CD31, CD34, CD45, CD73, CD90, CD105, CD166 and HLA-DR were analyzed by flow cytometry. The expression of embryonal markers Oct-4, Nanog, Sox-2, SSEA-1, SSEA-3, SSEA-4, TRA-1-60 and TRA 1-81 were studied by immunocytochemistry. The fC-MSC treated with specific induction medium were evaluated for their differentiation into (1) adipocytes and osteocytes (mesodermal cells) by Oil Red O and Alizarin Red staining, respectively, as well as by expression of lipoprotein lipase, PPARγ2 genes in adipocytes and osteopontin and RUNX2 genes in osteocytes by reverse-transcription polymerase chain reaction (RT-PCR); (2) neuronal (ectodermal) cells by expression of neuronal Filament-160 and Glial Fibrillar Acidic Protein by RT-PCR and immunocytochemistry; and (3) hepatocytic (endodermal) cells by expression of albumin by RT-PCR and immunocytochemistry, glycogen deposits by Periodic Acid Schiff staining and excretion of urea into the culture supernatant.
RESULTS
The fC-MSC expressed CD29, CD73, CD90, CD105, CD166 but lacked expression of CD31, CD34, CD45 and HLA-DR. They expressed embryonal markers, viz. Oct-4, Nanog, Sox-2, SSEA-1, SSEA-3, SSEA-4, TRA-1-81 but not TRA-1-60. On treatment with specific induction media, they differentiated into adipocytes and osteocytes, neuronal cells and hepatocytic cells.
CONCLUSION
Our results together suggest that fC-MSC are primitive stem cell types with a high degree of plasticity and, in addition to their suitability for cardiovascular regenerative therapy, they may have a wide spectrum of therapeutic applications in regenerative medicine.
目的
研究胎儿心脏间充质干细胞（fC-MSC）的胚胎标志物表达及其向所有胚层细胞的分化。
方法
从单个或多个胎儿心脏中分离出细胞，建立了 10 个独立的大鼠 fC-MSC 培养物，并在第 3、6、15 和 21 代进行了以下研究。通过流式细胞术分析表型标志物 CD29、CD31、CD34、CD45、CD73、CD90、CD105、CD166 和 HLA-DR。通过免疫细胞化学研究胚胎标志物 Oct-4、Nanog、Sox-2、SSEA-1、SSEA-3、SSEA-4、TRA-1-60 和 TRA 1-81 的表达。用特定诱导培养基处理 fC-MSC，通过油红 O 和茜素红染色分别评估其向脂肪细胞和成骨细胞（中胚层细胞）的分化，以及通过逆转录聚合酶链反应（RT-PCR）评估脂肪细胞中脂蛋白脂肪酶、PPARγ2 基因和骨细胞中骨桥蛋白和 RUNX2 基因的表达；通过 RT-PCR 和免疫细胞化学评估向神经细胞（外胚层）的分化；通过 RT-PCR 和免疫细胞化学评估向肝细胞（内胚层）的分化，通过 RT-PCR 评估白蛋白的表达，通过过碘酸希夫染色评估糖原沉积，通过向培养上清液中排泄尿素评估向肝细胞的分化。
结果
fC-MSC 表达 CD29、CD73、CD90、CD105、CD166，但缺乏 CD31、CD34、CD45 和 HLA-DR 的表达。它们表达胚胎标志物，即 Oct-4、Nanog、Sox-2、SSEA-1、SSEA-3、SSEA-4、TRA-1-81，但不表达 TRA-1-60。在特定诱导培养基的作用下，它们分化为脂肪细胞和成骨细胞、神经元细胞和肝细胞。
结论
我们的结果表明，fC-MSC 是具有高度可塑性的原始干细胞类型，除了适合心血管再生治疗外，它们在再生医学中可能具有广泛的治疗应用。
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