Fast but Durable Megakaryocyte Repopulation and Platelet Production in NOD/SCID Mice Transplanted with Ex-Vivo Expanded Human Cord Blood CD34 + Cells

Stefania Bruno, Monica Gunetti, Loretta Gammaitoni, Eliana Perissinotto, Luisa Caione, Fiorella Sanavio, Franca Fagioli, Massimo Aglietta, Wanda Piacibello

Research output: Contribution to journalArticle

Abstract

We have previously established a stroma-free culture with Flt-3 ligand (FL), stem cell factor (SCF), and thrombopoietin (TPO) that allows the maintenance and the expansion for several weeks of a cord blood (CB) CD34 + cell population capable of multilineage and long-lasting hematopoietic repopulation in non-obese diabetic/severe combined immunodeficient (NOD/SCID) mice. In this work the kinetics of megakarocyte (Mk)-engraftment that is often poor and delayed in CB transplantation, and human platelet (HuPlt) generation in NOD/SCID mice of baseline CD34+ cells (b34+), and of CD34+ cells reisolated after a 4-week expansion with FL+SCF+TPO (4w34+) were compared. With b34 + cells Mk-engraftment was first seen at week 3 (CD41+: 0.4%); 4w34+ cells allowed a more rapid Mk-engraftment (at weeks 2 and 3 the CD41+ cells were 0.3% and 0.8%). Circulating HuPlts were first seen at weeks 2 and 1, respectively. Mk-engraftment levels of b34 + and 4w34+ cells 6-8 weeks after transplantation were similar (12 ± 3.5 versus 15 ± 5% CD45+; 1.3 ± 0.5 versus 1.8 ± 0.5% CD41+ cells). Also serial transplant experiments were performed with expanded and reselected CB cells. In secondary and tertiary recipients the Mk population was detected with bone marrow fluorescence-activated cell sorter analysis; these experiments indicate the effective long-term repopulation of expanded cells. Selected CD34+ cells after a 4-week expansion with FL+SCF+TPO are more efficient in Mk engraftment than the same number of unmanipulated cells.

Original languageEnglish
Pages (from-to)135-143
Number of pages9
JournalStem Cells
Volume22
Issue number2
Publication statusPublished - 2004

Fingerprint

Megakaryocytes
SCID Mice
Fetal Blood
Blood Cells
Blood Platelets
Thrombopoietin
Stem Cell Factor
Transplantation
Population
Cell Count
Fluorescence
Bone Marrow
Maintenance
Transplants

Keywords

  • CD34 cell expansion
  • Cord blood
  • Megakaryocyte engraftment
  • NOD/SCID

ASJC Scopus subject areas

  • Cell Biology

Cite this

Fast but Durable Megakaryocyte Repopulation and Platelet Production in NOD/SCID Mice Transplanted with Ex-Vivo Expanded Human Cord Blood CD34 + Cells. / Bruno, Stefania; Gunetti, Monica; Gammaitoni, Loretta; Perissinotto, Eliana; Caione, Luisa; Sanavio, Fiorella; Fagioli, Franca; Aglietta, Massimo; Piacibello, Wanda.

In: Stem Cells, Vol. 22, No. 2, 2004, p. 135-143.

Research output: Contribution to journalArticle

Bruno, S, Gunetti, M, Gammaitoni, L, Perissinotto, E, Caione, L, Sanavio, F, Fagioli, F, Aglietta, M & Piacibello, W 2004, 'Fast but Durable Megakaryocyte Repopulation and Platelet Production in NOD/SCID Mice Transplanted with Ex-Vivo Expanded Human Cord Blood CD34 + Cells', Stem Cells, vol. 22, no. 2, pp. 135-143.
Bruno, Stefania ; Gunetti, Monica ; Gammaitoni, Loretta ; Perissinotto, Eliana ; Caione, Luisa ; Sanavio, Fiorella ; Fagioli, Franca ; Aglietta, Massimo ; Piacibello, Wanda. / Fast but Durable Megakaryocyte Repopulation and Platelet Production in NOD/SCID Mice Transplanted with Ex-Vivo Expanded Human Cord Blood CD34 + Cells. In: Stem Cells. 2004 ; Vol. 22, No. 2. pp. 135-143.
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AU - Gunetti, Monica

AU - Gammaitoni, Loretta

AU - Perissinotto, Eliana

AU - Caione, Luisa

AU - Sanavio, Fiorella

AU - Fagioli, Franca

AU - Aglietta, Massimo

AU - Piacibello, Wanda

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AB - We have previously established a stroma-free culture with Flt-3 ligand (FL), stem cell factor (SCF), and thrombopoietin (TPO) that allows the maintenance and the expansion for several weeks of a cord blood (CB) CD34 + cell population capable of multilineage and long-lasting hematopoietic repopulation in non-obese diabetic/severe combined immunodeficient (NOD/SCID) mice. In this work the kinetics of megakarocyte (Mk)-engraftment that is often poor and delayed in CB transplantation, and human platelet (HuPlt) generation in NOD/SCID mice of baseline CD34+ cells (b34+), and of CD34+ cells reisolated after a 4-week expansion with FL+SCF+TPO (4w34+) were compared. With b34 + cells Mk-engraftment was first seen at week 3 (CD41+: 0.4%); 4w34+ cells allowed a more rapid Mk-engraftment (at weeks 2 and 3 the CD41+ cells were 0.3% and 0.8%). Circulating HuPlts were first seen at weeks 2 and 1, respectively. Mk-engraftment levels of b34 + and 4w34+ cells 6-8 weeks after transplantation were similar (12 ± 3.5 versus 15 ± 5% CD45+; 1.3 ± 0.5 versus 1.8 ± 0.5% CD41+ cells). Also serial transplant experiments were performed with expanded and reselected CB cells. In secondary and tertiary recipients the Mk population was detected with bone marrow fluorescence-activated cell sorter analysis; these experiments indicate the effective long-term repopulation of expanded cells. Selected CD34+ cells after a 4-week expansion with FL+SCF+TPO are more efficient in Mk engraftment than the same number of unmanipulated cells.

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