X-Message-Number: 10737
Date: Tue, 10 Nov 1998 08:55:22 -0800
From: "Joseph J. Strout" <>
Subject: more on trehalose

I did a little lit search on trehalose and cryopreservation.  A few items
sound quite interesting...

Beattie, GM; Crowe, JH; Lopez, AD; Cirulli, V; Ricordi, C; Hayek,
A.  Trehalose: a cryoprotectant that enhances recovery and preserves
function of human pancreatic islets after long-term storage. Diabetes, 1997
Mar, 46(3):519-23.

From the abstract: We have developed a method to cryopreserve islets with
excellent survival of endocrine cells. Current methods use DMSO as
cryoprotectant. Our method involves introducing both DMSO and the
disaccharide trehalose into the cells during cooling. Uptake and release of
trehalose occurred during the thermotropic lipid-phase transition measured
in pancreatic endocrine cells between 5 degrees and 9 degrees C, using
[14C]trehalose. Recovery of adult islets after cryopreservation with 300
mmol/l trehalose was 92 vs. 58% using DMSO alone. In vitro function, in
terms of insulin content and release in response to secretagogues, was
indistinguishable from fresh islets. Grafts from islets cryopreserved with
trehalose contained 14-fold more insulin than grafts from islets
cryopreserved without trehalose. Results with human fetal islet-like cell
clusters (ICCs) were more pronounced: recovery from cryopreservation was
94%, compared with 42% without trehalose. ... Thus, the addition of
trehalose to cryopreservation protocols leads to previously unobtainable
survival rates of human pancreatic endocrine tissue.


Saha, S; Otoi, T; Takagi, M; Boediono, A; Sumantri, C; Suzuki, T.  Normal
calves obtained after direct transfer of vitrified bovine embryos using
ethylene glycol, trehalose, and polyvinylpyrrolidone.  Cryobiology, 1996
Jun, 33(3):291-9. (UI: 96260185)

From the abstract: ...embryos were vitrified using one of the three
treatments   (Treatment A, 40% ethylene glycol (EG); treatment B, 40% EG +
11.3%   trehalose; and treatment C, 40% EG + 11.3% trehalose + 20% PVP and
rehydrations) was performed directly in mPBS. Highest development   (84.1%)
and hatching rate (68.2%) were obtained when embryos were   vitrified with
the vitrification solution used in treatment C.


Yokomise, H; Inui, K; Wada, H; Ueda, M; Hitomi, S.  Long-term
cryopreservation can prevent rejection of canine tracheal allografts with
preservation of graft viability.  Journal of Thoracic and Cardiovascular
Surgery, 1996 May, 111(5):930-4.

From the abstract: Six to 10 rings of the trachea were removed from donor
dogs (n = 12), immersed in the preservative solution [containing
trehalose], and cryopreserved in a  deep freezer at -85 degrees C for 285
+/- 28 days (cryopreservation group).  Five rings of the mediastinal
trachea of recipient dogs were removed. The  cryopreserved tracheas were
thawed and transplanted to replace the excised mediastinal tracheas... The
anastomotic site  and graft were covered with an omental pedicle in both
groups. In the cryopreservation group, every animal, except one that was
killed for pathologic examination, survived more than 2 months. All the
grafts of this group were viable, and no stenosis or tracheomalacia was
observed.


These results suggest to me that trehalose may have value not just in
above-freezing preservation, but as a cryoprotectant for long-term storage
as well.  (And FWIW, several of the abstracts I read reported that
polyvinylpyrrolidone (PVP) may work even better in some cases, either alone
or in combination with trehalose.)

Speaking of such things, I was unable to make the 21CM seminar last Sunday.
Are any of the reports available?  Or any attendees who would care to
summarize?

Best regards,
-- Joe


,------------------------------------------------------------------.
|    Joseph J. Strout           Department of Neuroscience, UCSD   |
|                 http://www.strout.net              |
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