Functional and immunophenotypic characteristics of isolated CD105+ and fibroblast+ stromal cells from AML: implications for their plasticity along endothelial lineage

doi:10.1080/14653240310000092

Cytotherapy

Volume 5, Issue 1, 2003, Pages 66-79

https://doi.org/10.1080/14653240310000092 Get rights and content

Background

In vitro cultures of BM cells firm newly diagnosed patients with AML displayed a defective BM stromal compartment, with a reduced number offibroblast-colony-forming unit (CFU-F: 1 ± 1.25 SD) and a decreased proliferatrve ability. The purposes of our study were: 1) to select BM mesencbymal stem cells (MSC) and BM-derived stromal cells (BMDSCs) from AML patients at diagnosis and from healthy subjects, using an immunomagnetic system and either anti-CD105 or anti-fibroblast MAbs; 2) to study the immunophenotypic and functional properties of freshly isolated and cultured mesencbymal cells; 3) to test the in vitro plasticity of the selected cells to differentiate towards an endothelial phenotype.

Methods

Fresh mononuclear cells obtained from BM of 20 patients newly diagnosed with AML and from eight healthy subjects were selected by using anti-fibroblast and anti-CD105 MAbs. Freshly isolated cells were analyzed, characterized by flow cytometry using a wide panel of MAbs and seeded in long-term culture medium to assess CFU-F formation. The level of confluence after 30 days and functional capacity in a long-term colony-forming cell culture (LTC-CFC) were tested. Furthermore, the cultured selected cell populations were assayed for their ability to differentiate into an endothelial-like cell pbenotype with the addition of vascular endothelial growth factor (VEGF) and endothelial cell growth supplement (ECGS).

Results

In normal subjects the selection produced an increase of the CFU-F number of 2.6-fold with anti-fibroblast MAb and 2.1-fold with the anti-CDIOS MAb. Anti-fibroblast and anti-CD105 MAb selection from AML BM cells resulted in a statistically significant greater count ofCFU-F that was respectively 10.6-fold (P = 0.04) and 14.4-fold (P = 0.00001) higher in comparison with the unsdected AML samples. Interestingly, in 80% of AML samples immunoselection was also able to restore the capacity of the CFU-F to proliferate and form confluent stromal layers. The isolation of those layers sustained the proliferation and differentiation of bematopoietic stem cells in the LTC-CFC. The phenotypic profile of cultured BMDSCs was different from that of the freshly isolated cells, and changed in relation to the culture conditions: CD105⁺ selected cells cultured with VEGF and ECGS expressed endothelial markers, a finding that suggests that this cell subpopulation may have the potential to differentiate toward an endothelial-like phenotype.

Discussion

We report that immunomagnetic selection represents a valid tool for the selection of BM mesencbymal cells in samples obtained from both healthy subjects and patients with AML. This technique -was able to rescue two functional and immunophenotypic compartments related to two different selected populations. In particular, the CD105⁺ cells isolated in AML displayed, after stimulation with VEGF and ECGS, the ability to change towards an endothelial-like cell phenotype, thus revealing an unexpected plasticity. Both CD105⁺ and fibroblast⁺ cells once successfully isolated might represent sources of mesencbymal cells populations useful for in vitro investigations and, above all, as therapeutic devices.

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Functional and immunophenotypic characteristics of isolated CD105+ and fibroblast+ stromal cells from AML: implications for their plasticity along endothelial lineage

Background

Methods

Results

Discussion

Blood

Blood

Exp Hematol

Blood

Blood

Exp Hematol

Exp Neurol

Exp Hematol

Exp Hematol

Exp Hematol

Exp Hematol

Blood

Blood

Exp Hematol

Blood Cells Mol Dis

Cytotherapy

Blood

Bone

Exp Hematol

Biocbem Biopbys Res Commun

Stromal cell associated haematopoiesis

J Cell Physiol

Regulation of hematopoiesis by bone marrow stromal cells and their products

Annu Rev Immunol

Biology of the hematopoietic microenvironment

EurJ Haematol

Precursors for fibroblasts in different populations of hematopoietic cells as detected by the in vitro colony assay method

Exp Hematol

Factors required for bone marrow stromal fibroblast colony formation in vitro

Br J Haematol

Marrow stromal stem cells

J Cell Sci Suppl

Phenotypic and functional comparison of cultures of marrow-derived mesench- ymal stem cells (MSCs) and stromal cell

J Cell Pbysiol

Isolation and culture of endothelial cells from human bone marrow

Br J Haematol

Regulation of hematopoiesis by microvascular endothelium

Leuk Lymph

Stromal cells responsible for transferring the microenvironment of the hemopoietic tissues Cloning in vitro and retransplantation in vivo

Transplantation

Micro-SAGE analysis of a single human marrow stromal cell colony-forming unit- fibroblast

Cytotherapy

Propagation and senescence of human marrow stromal cells in culture: a simple colony-forming assay identifies samples with the greatest potential to propagate and differentiate

Br J Haematol

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J Hemat Stem Cell Res

Bone marrow mesenchymal cells: biological properties and clinical application

J Biol Regul Homeost Agents

Stem cell alchemy

NatMed

Stem cells have an identity crisis

Haematotberapy

Marrow stromal cells as stem cells for non hematopoietic tissues

Science

Multilineal potential of adult human mesenchymal stem cells

Science

Myogenic cells derived from rat bone marrow mesenchymal stem cells exposed to 5-azacytidine

Muscle Nerve

Muscle regeneration by bone marrow-derived myogenic progenitors

Science

Functional and immunophenotypic characteristics of isolated CD105⁺ and fibroblast⁺ stromal cells from AML: implications for their plasticity along endothelial lineage