This work was supported, in part, by a grant from the NIH (RO1 HL098228) to CKB.

Note: All animal procedures were approved by the Nationwide Children&#8217;s Hospital Institutional Animal Care and Use Committee.
1. Pulmonary Heart Valve Harvest from a Heart Valve Donor Mouse
<ol>
	<li>Autoclave all the surgical tools before the surgery: 1x fine scissors, 3x micro forceps, 2x micro vascular clamps, 1x clamp applying forceps, 1x micro needle holder, 1x spring scissors, 1x retractor.</li>
	<li>A 6-8 week old female C57BL/6 mouse is used as a pulmonary heart valve donor. Rem...

<ol>
	<li>Polito, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Increased+morbidity+and+mortality+in+very+preterm+VLBW+infants+with+congenital+heart+disease.">Increased morbidity and mortality in very preterm VLBW infants with congenital heart disease.</a> Intens Care Med. 39, 1104-1112 (2013).</li><li>Wren, C., Reinhardt, Z., Khawaja, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Twenty+year+trends+in+diagnosis+of+life+threatening+neonatal+cardiovascular+malformations.">Twenty year trends in diagnosis of life threatening neonatal cardiovascular malformations.</a> Arch Dis Child Fetal. 93, F33-F35 (2008).</li><li>Vacanti, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Beyond+transplantation+Third+annual+Samuel+Jason+Mixter+lecture.">Beyond transplantation Third annual Samuel Jason Mixter lecture.</a> Archives of surgery. 123, 545-549 (1960).</li><li>Tudorache, I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orthotopic+replacement+of+aortic+heart+valves+with+tissue-engineered+grafts.">Orthotopic replacement of aortic heart valves with tissue-engineered grafts.</a> Tissue engineering Part A. 19, 1686-1694 (2013).</li><li>van Geldorp, M. W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Patient+outcome+after+aortic+valve+replacement+with+a+mechanical+or+biological+prosthesis+weighing+lifetime+anticoagulant+related+event+risk+against+reoperation+risk.">Patient outcome after aortic valve replacement with a mechanical or biological prosthesis weighing lifetime anticoagulant related event risk against reoperation risk.</a> The Journal of thoracic and cardiovascular surgery. 137, 881-886 (2009).</li><li>El Oakley, R., Kleine, P., Bach, D. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Choice+of+prosthetic+heart+valve+in+today's+practice.">Choice of prosthetic heart valve in today's practice.</a> Circulation. 117, 253-256 (2008).</li><li>Quinn, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+Models+for+Bench+to+Bedside+Translation+of+Regenerative+Cardiac+Constructs.">Animal Models for Bench to Bedside Translation of Regenerative Cardiac Constructs.</a> Progress in Pediatric cardiology. 35, (2013).</li><li>Shinoka, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tissue+engineering+heart+valves+valve+leaflet+replacement+study+in+a+lamb+model.">Tissue engineering heart valves valve leaflet replacement study in a lamb model.</a> The Annals of thoracic surgery. 60, S513-S516 (1995).</li><li>Sodian, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tissue+engineering+of+heart+valves+in+vitro+experiences.">Tissue engineering of heart valves in vitro experiences.</a> The Annals of thoracic surgery. 70, 140-144 (2000).</li><li>Shinoka, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tissue+engineered+heart+valves.+Autologous+valve+leaflet+replacement+study+in+a+lamb+model.">Tissue engineered heart valves. Autologous valve leaflet replacement study in a lamb model.</a> Circulation. 94, II164-II168 (1996).</li><li>Sodian, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+biodegradable+three+dimensional+matrices+for+tissue+engineering+of+heart+valves.">Evaluation of biodegradable three dimensional matrices for tissue engineering of heart valves.</a> ASAIO journal (American Society for Artificial Internal Organs. 46, 107-110 (1992).</li><li>Sodian, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+in+vivo+experience+with+tissue-engineered+trileaflet+heart+valves).">Early in vivo experience with tissue-engineered trileaflet heart valves).</a> Circulation. 102, III22-III29 (2000).</li></ol>

The mortality rate of this procedure is close to 20%, which was mostly caused by hemorrhage at the PV transplantation site and anastomosis on the donor aorta to the recipient abdominal aorta. In most of the cases, the mortality rate decreases significantly 48 hr post surgery. The survival mice showed strong heart beats and blood flow through the implanted PV. The entire process takes four hours for an experienced micro surgeon. It will take roughly 250 mice to master the technique. The heterotopic heart transplantation i...

Congenital cardiovascular defects are one of the leading causes of infant mortality in the western world1,2.&#160; Among them, pulmonic valve stenosis and bicuspid aortic valve defects are a frequently occurring form3.&#160; Heart valve replacement surgery is a routine choice of reconstructive surgeries; however, complications including stenosis and calcification of the heart valve, and lifelong dependence on anticoagulants are a significant source of chronic ill health and death4-7. Moreover, the lack of growth potential requires revision surgeries, which further increases the mortality of those young patients4,8,9.
In an attempt to develop a functional replacement heart valve with growth potential, Shinoka et al. seeded autologous cells onto a biodegradable synthetic heart valve8. The synthetic valve transformed to a native heart valve like structure with growth potential. Preliminary large animal studies demonstrated the feasibility of using this methodology to create a functional heart valve10.&#160; However, long term implantation studies demonstrated poor durability due to progressive thickening of the valve neotissue resulting in narrowing of the heart valve. Work from Sodian et al. used the Shinoka methodology, but ultimately replaced the PGA matrix with a biodegradable elastomer, which gave the biomechanical properties of the tissue engineered valve construct a more physiological profile9,11,12. In the in vivo study, despite the success of the implantation, a confluent endothelial cell lining was not formed which could limit the long term success of this scaffold12.
In order to rationally design an improved second generation synthetic heart valve, a murine model of heart valve transplantation was created to investigate the cellular and molecular mechanisms underlying neotissue formation, valve thickening, and stenosis development. Murine models offer a vast array of molecular reagents, including transgenics, which are not readily available in other species7. In this heart valve transplantation model, an ex vivo syngeneic pulmonary heart valve replacement was performed first; and then the heart with the implanted heart valve was implanted heterotopically into a syngeneic host using a microsurgical technique. This model enables heart valve replacement without the need for cardiopulmonary bypass.
In this paper, a detailed explanation of a heart valve harvest, donor heart preparations, heart valve transplantation, and heterotopic heart transplantation is described. The results showed a continuous heartbeat from the donor heart, which was independent of the recipient heartbeat. The blood flow through the implanted pulmonary valve was measured using a high frequency ultrasound system with a pulsed wave Doppler.

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transplantation of pulmonary valve using a mouse model of heterotopic heart transplantation

Tissue engineered heart valves, especially decellularized valves, are starting to gain momentum in clinical use of reconstructive surgery with mixed results. However, the cellular and molecular mechanisms of the neotissue development, valve thickening, and stenosis development are not researched extensively. To answer the above questions, we developed a murine heterotopic heart valve transplantation model. A heart valve was harvested from a valve donor mouse and transplanted to a heart donor mouse. The heart with a new valve was transplanted heterotopically to a recipient mouse. The transplanted heart showed its own heartbeat, independent of the recipient&#8217;s heartbeat. The blood flow was quantified using a high frequency ultrasound system with a pulsed wave Doppler. The flow through the implanted pulmonary valve showed forward flow with minimal regurgitation and the peak flow was close to 100 mm/sec. This murine model of heart valve transplantation is highly versatile, so it can be modified and adapted to provide different hemodynamic environments and/or can be used with various transgenic mice to study neotissue development in a tissue engineered heart valve.

Figure 1 illustrates the schematics of the heart valve transplantation model using heterotopic heart transplantation. The heart valve was harvested from a donor heart and implanted onto a heart from a second donor mouse. Then the heart with the new heart valve was implanted to the abdomen of a recipient mouse. Figure 2 shows an illustration of the implanted heart on the abdominal space (A), right after heart transplantation (B), and 5 min after transplan...

Watch this Scientific Journal Video about Transplantation of Pulmonary Valve Using a Mouse Model of Heterotopic Heart Transplantation at JoVE.com

Transplantation of Pulmonary Valve Using a Mouse Model of Heterotopic Heart Transplantation

In order to understand the cellular and molecular mechanisms underlying neotissue formation and stenosis development in tissue engineered heart valves, a murine model of heterotopic heart valve transplantation was developed. A pulmonary heart valve was transplanted to recipient using the heterotopic heart transplantation technique.

Tissue engineered heart valves, especially decellularized valves, are starting to gain momentum in clinical use of reconstructive surgery with mixed ...

transplantation-pulmonary-valve-using-mouse-model-heterotopic-heart

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Medicine

18.7K vistas.  Nationwide Children's Hospital.  Con el fin de comprender los mecanismos celulares y moleculares que subyacen a la formación de neotejido y el desarrollo de estenosis en las válvulas del corazón de la ingeniería tisular, se desarrolló un modelo murino de trasplante heterotópico de válvula cardiaca. Una válvula cardíaca pulmonar fue trasplantado al receptor mediante la técnica de trasplante cardíaco heterotópico. 