placental digestion

Heidi Engelhardt hengelha at uoguelph.ca
Thu Sep 28 11:54:18 EST 2000


Bill Hyun wrote:
>
> Hello,
> Does anyone have a protocol or reference for digesting mouse or human
> placenta to single cells for sorting?

Hi Bill.  There are lots of people doing this in both mouse, human, and
other species.  With human, one usually gets first trimester (ie.
abortions) or term placenta.  A sampler:

Abadia-Molina et al 1996 Am J Reprod Immunol 31:109-123
(human/first trimester/collagenase digest)

King et al 1993 Dev Immunol 1:169-190
(human/first trimester/DECIDUA - mechanical sieving plus gradient)

(PIs on both of above are Ashley King and YW Loke)

Ritson and Bulmer 1987 J Immunol Methods 104:231-36
(human/decidua but good - compares effcts of various methods on
viability, leukocyte content, etc)

Stewart and Mukhtar 1988 Placenta 9:417-25
(mouse/enzymatic)

Zuckerman and Head 1988 Placenta 7:349-
(mouse/dispase plus gradient)

We use a collagenase digestion (plus a pinch of DNase) followed by
gradient centrifugation to harvest leukocytes from pig endometrium.  A
similar approach works with pig trophoblast (Biol Reprod 1994
51:1279-84).

I would advise staying away from dispase, protease and maybe even
trypsin if you want to assess surface markers immediately (ie. without
culturing first).

Heidi

--
**  note new email address  **

Heidi Engelhardt, Adjunct Professor
Department of Animal & Poultry Science
University of Guelph
Guelph, Ontario
CANADA N1G 2W1

email:	hengelha at uoguelph.ca
phone:	(519) 824-4120, ext. 3658
fax:	(519) 767-0573



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