In Vitro Grown Micro-Tissues for Cardiac Cell Replacement Therapy in Vivo.
In Vitro Grown Micro-Tissues for Cardiac Cell Replacement Therapy in Vivo.
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DOI：
10.33594/000000092
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发表时间：
2019-01-01
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影响因子：
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通讯作者：
Pfannkuche, Kurt
中科院分区：
文献类型：
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作者：
Sahito, Raja Ghazanfar Ali;Sheng, Xiaowu;Pfannkuche, Kurt
背景/目的：不同的方法被认为是改善心脏重建医学的方法，在这种情况下，直接输送多能干细胞来源的心肌细胞(PSC-CMS)似乎是非常有前途的。然而，移植后细胞持续性低仍然是阻碍这一方法的瓶颈。在此，我们提出了一种新的策略来克服PSC-CMS在移植后早期低植入到健康和冷冻损伤同基因小鼠的心肌中。方法：成年小鼠骨髓间充质干细胞(MSCs)和PSC-CMS在温度响应性聚合物上共培养，然后通过降温分离，导致由这两种细胞组成的细胞团(微组织)的无酶生成。显微组织被移植到健康和冷冻损伤的小鼠心脏中。用实时荧光定量聚合酶链式反应(Real-Time-PCR)测定细胞短期存留率。生物发光成像进行细胞纵向示踪，连续4周。结果：由PSC-CMS和MSCs组成的体外培养的微组织可使移植后1天的心肌细胞存留量增加10倍，但不能完全挽救移植后1~2天的进一步细胞丢失。在健康心脏组织中可检测到中性粒细胞渗入，这可能是由于细胞植入而不是移植导管对组织的损伤。通过生物发光成像对注射的PSC-CMS进行跟踪，并成功检测了长达四周的PSC-CMS。结论：该方法表明，体外培养的微组织可能有助于心肌细胞替代疗法的发展。
BACKGROUND/AIMS: Different approaches have been considered to improve heart reconstructive medicine and direct delivery of pluripotent stem cell-derived cardiomyocytes (PSC-CMs) appears to be highly promising in this context. However, low cell persistence post-transplantation remains a bottleneck hindering the approach. Here, we present a novel strategy to overcome the low engraftment of PSC-CMs during the early post-transplantation phase into the myocardium of both healthy and cryoinjured syngeneic mice.METHODS: Adult murine bone marrow mesenchymal stem cells (MSCs) and PSC-CMs were co-cultured on thermo-responsive polymers and later detached through temperature reduction, resulting in the protease-free generation of cell clusters (micro-tissues) composed of both cells types. Micro-tissues were transplanted into healthy and cryo-injured murine hearts. Short term cell retention was quantified by real-time-PCR. Longitudinal cell tracking was performed by bioluminescence imaging for four weeks. Transplanted cells were further detected by immunofluorescence staining of tissue sections.RESULTS: We demonstrated that in vitro grown micro-tissues consisting of PSC-CMs and MSCs can increase cardiomyocyte retention by >10fold one day post-transplantation, but could not fully rescue a further cell loss between day 1 and day 2. Neutrophil infiltration into the transplanted area was detected in healthy hearts and could be attributed to the cellular implantation rather than tissue damage exerted by the transplantation cannula. Injected PSC-CMs were tracked and successfully detected for up to four weeks by bioluminescence imaging.CONCLUSION: This approach demonstrated that in vitro grown micro-tissues might contribute to the development of cardiac cell replacement therapies.