X-Message-Number: 31028
Date: Mon, 8 Sep 2008 09:15:56 -0700 (PDT)
From:
Subject: Cryopreservation of whole murine and porcine livers.
[Looks worth checking out.]
Rejuvenation Res. 2008 Aug;11(4):765-72.
Cryopreservation of whole murine and porcine livers.
Gavish Z, Ben-Haim M, Arav A. Core Dynamics Ltd., Nes Ziona, Israel.,
Department of Animal Science, ARO, Bet Pagan, Israel.
Abstract Preservation of vascularized organs, such as the liver, is limited
to 24 h before destructive processes disqualify them for transplantation.
This narrow window of opportunity prevents the performance of optimal
pathogen screening and matching tests and possibly results in the need for
retransplantation. Numerous problems are associated with freezing and
thawing a whole liver while preserving its viability upon thawing, including
complicated geometry, poor heat transfer, release of latent heat, and the
difficulty of generating a uniform cooling rate. On the basis of our past
success with sheep ovaries, we have now applied our novel freezing technique
to a larger solid organ, the liver. Whole rat and pig livers were frozen
and thawed using directional solidification apparatus, and viability of
these livers was tested by means of integrity and functionality in vitro and
in auxiliary liver transplantation. The thawed rat and porcine livers were
intact and demonstrated >80% viability. Histology revealed normal
architecture. Bile production and blood flow following auxiliary
transplantation were normal as well. Our encouraging results in applying
this novel cryopreservation technique in rat and pig livers suggest that
this method may enable better human organ donor-recipient matching in the
future.
PMID: 18729808
J Thorac Cardiovasc Surg. 2008 Mar;135(3):666-72, 672.e1.
Successful restoration of function of frozen and thawed isolated rat hearts.
Elami A, Gavish Z, Korach A, Houminer E, Schneider A, Schwalb H, Arav A.The
Department of Cardiothoracic Surgery and the Joseph Lunenfeld Cardiac
Surgery Research Center, Hadassah-Hebrew University Medical Center,
Jerusalem, Israel.
OBJECTIVE: Long-term organ preservation for transplantation may allow
optimal donor-recipient matching with potential reduction in the incidence
and severity of rejection. Complete cessation of metabolism may be obtained
by freezing. Previous attempts to freeze intact mammalian hearts were
limited to -3.6 degrees C, restricting tissue ice content to 34%. We
hypothesized that our method will allow recovery of function of the intact
rat heart after freezing to -8 degrees C, a temperature at which most of the
tissue water is frozen. METHODS: Isolated rat hearts were attached to a
Langendorff apparatus. After normothermic perfusion, cold cardioplegia was
induced followed by perfusion with a cryoprotecting agent. Hearts were than
frozen to -8 degrees C (45 +/- 8 minutes), thawed, and reperfused (60
minutes). RESULTS: All frozen and thawed hearts regained normal electric
activity. At -8 degrees C, ice content was 64.36% +/- 13%. The use of 10%
ethylene glycol for cryoprotection (n = 13) resulted in recovery (mean +/-
standard deviation) of 49.7% +/- 21.8% of +dP/dt, 48.0% +/- 23.5% of -dP/dt,
65.2% +/- 30.8% of coronary flow, and 50.4% +/- 23.9% of left ventricular
developed pressure. Hearts in this group (n = 4) maintained 81.3% +/- 10%
viability compared with 69.3% +/- 14% (not significant) in control hearts
kept at 0 degrees C for the same duration. Energy stores, represented by
adenosine triphosphate and phosphocreatine, were depleted to 12.2 +/- 6.1
micromol/g dry weight and 22.5 +/- 6.4 micromol/g dry weight, respectively,
compared with 19.0 +/- 2.5 micromol/g dry weight and 36.6 +/- 3.0 micromol/g
dry weight, respectively (P < .05) in the control hearts. The integrity of
muscle fibers and intracellular organelles after thawing and reperfusion was
demonstrated by electron microscopy. CONCLUSION: We demonstrate for the
first time the feasibility of functional recovery after freezing and thawing
of the isolated rat heart while maintaining structural integrity and
viability.
PMID: 18329491
Hum Reprod. 2005 Dec;20(12):3554-9. Epub 2005 Sep 20.
Oocyte recovery, embryo development and ovarian function after cryopreservation
and transplantation of whole sheep ovary.
Arav A, Revel A, Nathan Y, Bor A, Gacitua H, Yavin S, Gavish Z, Uri M, Elami
A. Institute of Animal Science, Agricultural Research Organization (ARO),
the Volcani Center, P.O.B. 6, Bet Dagan 50250, Israel.
BACKGROUND: Successful cryopreservation of a whole ovary may provide a
solution for women with premature ovarian failure. The aim of this study was
to evaluate the function of cryopreserved whole sheep ovaries both in vitro
and in vivo. METHODS: Transplantation of frozen-thawed intact ovaries was
performed on eight sheep by artery and vein anastomosis to the contralateral
ovarian artery and vein. The remaining ovary was removed. Oocyte aspiration
was performed 1 and 4 months post-transplantation. Serum progesterone
levels were measured after 24 and 36 months. Magnetic resonance imaging
(MRI) was carried out 12 months after transplantation. RESULTS: Progesterone
activity was detected in three sheep from 24 to 36 months
post-transplantation. Oocyte retrieval was successful in two sheep and
parthenogenic activation has resulted in embryonic development up to the
8-cell stage. MRI revealed an intact ovary with small follicles and intact
blood vessels. CONCLUSIONS: Whole ovaries, and the follicles and blood
vessels they contain, are able to survive cryopreservation. In addition, MRI
has shown that blood vessels were intact and that normal blood flow had
resumed to the transplant. We conclude that immediate and long-term hormonal
restoration and normal ovulation is possible after cryopreservation and
transplantation of whole ovaries in sheep.
PMID: 16174650
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