Volume 21 Issue 4
Apr.  2023
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Article Navigation >  Chinese Journal of General Practice  > 2023  >  21(4): 593-597
SUN Zhigang, XIE Ying, LI Haihui, SHEN Guozheng, LIU Xuegang. An acellular protocol of whole organ perfusion in rat heart and evaluation of its acellular effect[J]. Chinese Journal of General Practice, 2023, 21(4): 593-597. doi: 10.16766/j.cnki.issn.1674-4152.002938
Citation: SUN Zhigang, XIE Ying, LI Haihui, SHEN Guozheng, LIU Xuegang. An acellular protocol of whole organ perfusion in rat heart and evaluation of its acellular effect[J]. Chinese Journal of General Practice, 2023, 21(4): 593-597. doi: 10.16766/j.cnki.issn.1674-4152.002938
shu

An acellular protocol of whole organ perfusion in rat heart and evaluation of its acellular effect

doi: 10.16766/j.cnki.issn.1674-4152.002938
  • 1.

    Graduate School, Bengbu Medical College, Bengbu, Anhui 233000, China

Funds:

 KJ2019A0362

  • Received Date: 2022-11-15
    Available Online: 2023-05-31
    Abstract
  •   Objective  A new perfusion decellularization protocol was used to decellularize isolated SD rat hearts, and the decellularization effect was tested and evaluated.  Methods  Twelve 2-month-old male SD rats from the same litter were randomly divided into decellularized and control groups according to the random number table method, with 6 rats in each group. The hearts of each rat were removed after anesthesia and systemic heparinization. The hearts of the decellularized group were weighed, and then the perfusion needle was inserted retrograde into the ascending aorta and the perfusion device was connected for decellularization, and the changes of heart appearance were observed during the decellularization process. A small amount of tissue was taken from the same part of the left auricle, right auricle, apical region and right ventricular outflow tract of each heart to determine the dsDNA content, and the remaining tissue was prepared in paraffin sections and stained with HE and Masson stain, and then images were collected for analysis.  Results  During decellularization, the heart color gradually became light red, then became relatively transparent, and finally became milky white, and the coronary vascular structure was well preserved as seen by the naked eye. The heart mass before and after decellularization in the decellularized group were (1.27±0.03) g and (0.49±0.02) g, respectively, with statistically significant differences (t=111.164, P < 0.01). The differences in dsDNA content measured at each site in the hearts of both groups were statistically significant (all P < 0.01), and the dsDNA content measured at each site in the decellularized group was significantly lower than 50 ng/mg. Typical myocardial tissue sections were seen in HE-stained sections and Masson stained sections of the control group. In contrast, HE-stained sections and Masson stained sections of the decellularized group were reticulated, with no obvious cellular or nuclear structures, and the coronary artery contours were visible within them.  Conclusion  This decellularization protocol is simple and reliable, and is a more ideal solution for the preparation of cardiac whole-organ decellularized biological scaffolds.

     

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