Contribution of bone marrow-derived hematopoietic stem/progenitor cells to the generation of donor-marker⁺ cardiomyocytes in vivo.
Contribution of bone marrow-derived hematopoietic stem/progenitor cells to the generation of donor-marker⁺ cardiomyocytes in vivo.
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DOI：
10.1371/journal.pone.0062506
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Akashi K
中科院分区：
文献类型：
--
作者：
Fukata M;Ishikawa F;Najima Y;Yamauchi T;Saito Y;Takenaka K;Miyawaki K;Shimazu H;Shimoda K;Kanemaru T;Nakamura K;Odashiro K;Nagafuji K;Harada M;Akashi K
明确鉴定与心肌发生相关的细胞类型和机制对于有效的心脏再生医学至关重要。我们的目的是鉴定可以产生心肌细胞的细胞群，并澄清供体标记物+心肌细胞的产生是否需要BM衍生细胞和受体心肌细胞之间的细胞融合。将来自绿色荧光蛋白（GFP）小鼠的纯化的BM干/祖细胞移植到C57 BL/6小鼠或青色荧光蛋白（CFP）转基因小鼠中。从脐带血中分离纯化人造血干细胞（HSCs），移植到免疫功能低下的NOD/SCID/IL 2 r γ敲除小鼠体内。在免疫荧光染色后，通过共聚焦显微镜分析心脏组织中的GFP+细胞的心肌细胞的抗原性。GFP+供体来源的细胞、GFP+CFP+融合细胞和CFP+宿主来源的细胞通过线性解混分析来区分。通过实时定量聚合酶链反应评价异种受体心脏的人心肌细胞基因的表达。在C57 BL/6受体中，Lin−/lowCD 45+造血细胞比Lin−/lowCD 45 −间充质细胞产生更多的GFP+心肌细胞（每个受体37.0+/-23.9 vs 0.00+/-0.00 GFP+心肌细胞，P = 0.0095）。 移植的纯化HSC（Lin−/lowSca-1+或Lin−Sca-1+c-Kit+或CD 34 −Lin−Sca-1+c-Kit+）的数量与GFP+心肌细胞的数量相关（每个细胞组分中P<0.05），并且在CD 34 −Lin−Sca-1+c-Kit+受体中，每注射细胞剂量的GFP+心肌细胞的发生率最高。在造血祖细胞中，总髓样祖细胞产生的GFP+心肌细胞数量多于普通淋巴样祖细胞（每个受体12.8+/-10.7 vs 0.67+/-1.00 GFP+心肌细胞，P = 0.0021）。 在CFP受体中，所有GFP+心肌细胞检测共表达CFP。人肌钙蛋白C和肌球蛋白重链6的转录本中检测到的一些异种受体的心脏组织。我们的研究结果表明，造血干细胞通过骨髓中间体融合依赖机制导致心肌细胞的产生。使用骨髓衍生物作为供体细胞可能会允许更有效的基于细胞的心脏修复治疗。
Definite identification of the cell types and the mechanism relevant to cardiomyogenesis is essential for effective cardiac regenerative medicine. We aimed to identify the cell populations that can generate cardiomyocytes and to clarify whether generation of donor-marker+ cardiomyocytes requires cell fusion between BM-derived cells and recipient cardiomyocytes. Purified BM stem/progenitor cells from green fluorescence protein (GFP) mice were transplanted into C57BL/6 mice or cyan fluorescence protein (CFP)-transgenic mice. Purified human hematopoietic stem cells (HSCs) from cord blood were transplanted into immune-compromised NOD/SCID/IL2rγnull mice. GFP+ cells in the cardiac tissue were analyzed for the antigenecity of a cardiomyocyte by confocal microscopy following immunofluorescence staining. GFP+ donor-derived cells, GFP+CFP+ fused cells, and CFP+ recipient-derived cells were distinguished by linear unmixing analysis. Hearts of xenogeneic recipients were evaluated for the expression of human cardiomyocyte genes by real-time quantitative polymerase chain reaction. In C57BL/6 recipients, Lin−/lowCD45+ hematopoietic cells generated greater number of GFP+ cardiomyocytes than Lin−/lowCD45− mesenchymal cells (37.0+/−23.9 vs 0.00+/−0.00 GFP+ cardiomyocytes per a recipient, P = 0.0095). The number of transplanted purified HSCs (Lin−/lowSca-1+ or Lin−Sca-1+c-Kit+ or CD34−Lin−Sca-1+c-Kit+) showed correlation to the number of GFP+ cardiomyocytes (P<0.05 in each cell fraction), and the incidence of GFP+ cardiomyocytes per injected cell dose was greatest in CD34−Lin−Sca-1+c-Kit+ recipients. Of the hematopoietic progenitors, total myeloid progenitors generated greater number of GFP+ cardiomyocytes than common lymphoid progenitors (12.8+/−10.7 vs 0.67+/−1.00 GFP+ cardiomyocytes per a recipient, P = 0.0021). In CFP recipients, all GFP+ cardiomyocytes examined coexpressed CFP. Human troponin C and myosin heavy chain 6 transcripts were detected in the cardiac tissue of some of the xenogeneic recipients. Our results indicate that HSCs resulted in the generation of cardiomyocytes via myeloid intermediates by fusion-dependent mechanism. The use of myeloid derivatives as donor cells could potentially allow more effective cell-based therapy for cardiac repair.
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影响因子：
20.1
作者：
Ferreira-Martins J;Ogórek B;Cappetta D;Matsuda A;Signore S;D'Amario D;Kostyla J;Steadman E;Ide-Iwata N;Sanada F;Iaffaldano G;Ottolenghi S;Hosoda T;Leri A;Kajstura J;Anversa P;Rota M
通讯作者：
Rota M
影响因子：
64.8
作者：
Alvarez-Dolado, M;Pardal, R;Alvarez-Buylla, A
通讯作者：
Alvarez-Buylla, A
影响因子：
20.1
作者：
Kajstura, J;Rota, M;Anversa, P
通讯作者：
Anversa, P
影响因子：
64.8
作者：
Conboy, IM;Conboy, MJ;Rando, TA
通讯作者：
Rando, TA
影响因子：
20.3
作者：
Kawada, H;Fujita, J;Fukuda, K
通讯作者：
Fukuda, K