Sahito Raja Ghazanfar Ali, Sheng Xiaowu, Maass Martina, Mikhael Nelly, Hamad Sarkawt, Heras-Bautista Carlos O, Derichsweiler Daniel, Spitkovsky Dimitry, Suhr Frank, Khalil Markus, Brockmeier Konrad, Halbach Marcel, Saric Tomo, Hescheler Jürgen, Krausgrill Benjamin, Pfannkuche Kurt
Center for Physiology and Pathophysiology, Institute of Neurophysiology, Medical Faculty, University of Cologne, Cologne, Germany.
Hunan Cancer Hospital and Affiliated Cancer Hospital of Xiangya School of Medicine, Central South University, Changsha, China.
Cell Physiol Biochem. 2019;52(6):1309-1324. doi: 10.33594/000000092.
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.
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.
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.
This approach demonstrated that in vitro grown micro-tissues might contribute to the development of cardiac cell replacement therapies.
背景/目的：人们已经考虑了不同的方法来改进心脏重建医学，在这种情况下，直接递送多能干细胞衍生的心肌细胞（PSC-CMs）似乎非常有前景。然而，移植后细胞持久性低仍然是阻碍该方法的一个瓶颈。在这里，我们提出了一种新策略，以克服PSC-CMs在移植后早期阶段植入健康和冷冻损伤的同基因小鼠心肌中的低植入率问题。
将成年小鼠骨髓间充质干细胞（MSCs）和PSC-CMs在热响应聚合物上共培养，随后通过降温使其分离，从而在无蛋白酶的情况下生成由两种细胞类型组成的细胞簇（微组织）。将微组织移植到健康和冷冻损伤的小鼠心脏中。通过实时PCR对短期细胞保留情况进行定量。通过生物发光成像进行为期四周的纵向细胞追踪。通过组织切片的免疫荧光染色进一步检测移植细胞。
我们证明，由PSC-CMs和MSCs组成的体外培养微组织在移植后一天可使心肌细胞保留率提高10倍以上，但无法完全挽救第1天至第2天之间进一步的细胞损失。在健康心脏中检测到中性粒细胞浸润到移植区域，这可能归因于细胞植入而非移植套管造成的组织损伤。通过生物发光成像对注射的PSC-CMs进行追踪，并成功检测长达四周。
该方法表明，体外培养的微组织可能有助于心脏细胞替代疗法的发展。
