Use of CD9 expression to enrich for porcine hematopoietic progenitors.

Abstract:

OBJECTIVE:The aim of this study was to develop novel markers for enrichment of hematopoietic progenitors from bone marrow of swine. MATERIALS AND METHODS:We previously showed that pig bone marrow contains a "side population" (SP) of Hoechst dye-effluxing cells that resembles the hematopoietic stem cell (HSC)-containing murine SP and therefore represents a putative pig stem cell population. We screened a panel of monoclonal antibodies for those that allowed positive or negative enrichment of porcine SP cells and tested one of these for enrichment of hematopoietic progenitors in short-term and long-term in vitro assays. We then screened an expression library to clone the gene whose product is recognized by this antibody. RESULTS:Among a panel of 35 monoclonal lines screened, we found three that were useful for positive enrichment of SP cells and seven for negative enrichment. The 4-6 monoclonal line, allowing around 10-fold negative enrichment of SP cells, recognized the product of the porcine CD9 gene. Hematopoietic progenitors measured by short-term colony-forming unit and long-term cobblestone area-forming cell assays were around 10-fold enriched in the CD9(negative/low) fraction and were significantly depleted in the CD9(high) fraction. CONCLUSIONS:The antibody against the porcine CD9 gene product may be of use for enrichment of porcine hematopoietic stem cells. This approach to identify novel markers for enrichment of hematopoietic progenitors may be applicable to other mammalian species.

journal_name

Exp Hematol

journal_title

Experimental hematology

authors

Heinz M,Huang CA,Emery DW,Giovino MA,LeGuern A,Kurilla-Mahon B,Theodore P,Arn JS,Sykes M,Mulligan R,Down JD,Sachs DH,Goodell MA

doi

10.1016/s0301-472x(02)00835-4

keywords:

subject

Has Abstract

pub_date

2002-07-01 00:00:00

pages

809-15

issue

7

eissn

0301-472X

issn

1873-2399

pii

S0301472X02008354

journal_volume

30

pub_type

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