Long-term bovine hematopoietic engraftment with clone-derived stem cells.
Therapeutic cloning by somatic cell nuclear transfer offers potential for treatment of a wide range of degenerative disease. Nuclear transplantation with neo (r)-marked somatic nuclei from 10-13-year-old cows was used to generate cloned bovine fetuses. Clone fetal liver (FL) hematopoietic stem cells (HSC) were transplanted into two busulfan-treated and one untreated nuclear donor cows. Hematopoiesis was monitored over 13-16 months by in vitro progenitor and HSC assays. Chimerism was demonstrated by PCR in blood, marrow, lymph nodes, and endothelium, peaking at levels of 9-17% in blood granulocytes but at lower levels in lymphocyte subsets (0.1-0.01%). Circulating progenitors showed high levels of chimerism (up to 60% neo (r+)) with persisting fetal features. At sacrifice, the animal that had no pre-transplant myelosupression showed persisting donor cells in blood and lymph nodes, and in marrow 0.25% of progenitor cells and a detectable fraction of stem cells were neo (r+). The fetal HSC showed a 10-fold competition advantage over adult HSC. Cloning generated histocompatible HSC capable of long-term multilineage engraftment in a large animal model.
Lanza
Robert
R
Advanced Cell Technology, Worcester, Massachusetts, USA.
Shieh
J-H
JH
Wettstein
Peter J
PJ
Sweeney
Raymond W
RW
Wu
Kaida
K
Weisz
Anat
A
Borson
Nancy
N
Henderson
Boyd
B
West
Michael D
MD
Moore
Malcolm A S
MA
D000818:Animals; D001483:Base Sequence; D001854:Bone Marrow Cells / Q000166:cytology; D002417:Cattle; D002478:Cells, Cultured; D002999:Clone Cells; D017931:DNA Primers; D004730:Endothelium, Vascular / Q000166:cytology; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D008099:Liver / Q000166:cytology / Q000196:embryology; D008198:Lymph Nodes / Q000166:cytology; D016133:Polymerase Chain Reaction; D033581:Stem Cell Transplantation
D017931:DNA Primers