Kinetics of hematopoietic stem cells and supportive activities of stromal cells in a three-dimensional bone marrow culture system

Graft failure is a mortal complication in allogeneic bone marrow transplantation (BMT); T cells and natural killer cells are responsible for graft rejection. However, we have recently demonstrated that the recruitment of donor-derived stromal cells prevents graft failure in allogeneic BMT. This finding prompted us to examine whether a major histocompatibility complex (MHC) restriction exists between hematopoietic stem cells (HSCs) and stromal cells. We transplanted bone marrow cells (BMCs) and bones obtained from various mouse strains and analyzed the cells that accumulated in the engrafted bones. Statistically significant cell accumulation was found in the engrafted bone, which had the same H-2 phenotype as that of the BMCs, whereas only few cells were detected in the engrafted bones of the third-party H-2 phenotypes during the 4 to 6 weeks after BMT. Moreover, the BMCs obtained from the MHC-compatible bone showed significant numbers of both colony-forming units in culture (CFU-C) and spleen colony-forming units (CFU-S). These findings strongly suggest that an MHC restriction exists between HSCs and stromal cells.

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Major Histocompatibility Complex Restriction Between Hematopoietic Stem Cells and Stromal Cells In Vivo

Blood ◽

10.1182/blood.v89.1.49 ◽

1997 ◽

Vol 89 (1) ◽

pp. 49-54 ◽

Cited By ~ 85

Author(s):

Futoshi Hashimoto ◽

Kikuya Sugiura ◽

Kyoichi Inoue ◽

Susumu Ikehara

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Major Histocompatibility Complex ◽

Hematopoietic Stem Cells ◽

Stromal Cells ◽

Graft Failure ◽

Major Histocompatibility ◽

Hematopoietic Stem ◽

Histocompatibility Complex ◽

Colony Forming Units

Abstract Graft failure is a mortal complication in allogeneic bone marrow transplantation (BMT); T cells and natural killer cells are responsible for graft rejection. However, we have recently demonstrated that the recruitment of donor-derived stromal cells prevents graft failure in allogeneic BMT. This finding prompted us to examine whether a major histocompatibility complex (MHC) restriction exists between hematopoietic stem cells (HSCs) and stromal cells. We transplanted bone marrow cells (BMCs) and bones obtained from various mouse strains and analyzed the cells that accumulated in the engrafted bones. Statistically significant cell accumulation was found in the engrafted bone, which had the same H-2 phenotype as that of the BMCs, whereas only few cells were detected in the engrafted bones of the third-party H-2 phenotypes during the 4 to 6 weeks after BMT. Moreover, the BMCs obtained from the MHC-compatible bone showed significant numbers of both colony-forming units in culture (CFU-C) and spleen colony-forming units (CFU-S). These findings strongly suggest that an MHC restriction exists between HSCs and stromal cells.

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Induction of Adipocyte Differentiation and Reduction of Hematopoesis-Supporting Activity of Mouse Stromal Cells by Suppression of Transcription Factor GATA-2.

Blood ◽

10.1182/blood.v106.11.1392.1392 ◽

2005 ◽

Vol 106 (11) ◽

pp. 1392-1392

Author(s):

Yoko Okitsu ◽

Hideo Harigae ◽

Masanori Seki ◽

Toru Fujiwara ◽

Shinichiro Takahashi ◽

...

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Transcription Factor ◽

Hematopoietic Stem Cells ◽

Stromal Cells ◽

Colony Formation ◽

Adipocyte Differentiation ◽

Fetal Liver ◽

Expression Level ◽

Hematopoietic Stem

Abstract (Introduction) Aplastic anemia (AA) is characterized by peripheral pancytopenia and fatty bone marrow. An immunological attack to hematopoietic stem cells has been thought to be responsible for the development of the disease. Previously, we reported the expression of transcription factor GATA-2 is significantly decreased in CD34 positive cells in AA. Together with the phenotypes of hematopoietic stem cells in GATA-2 hetero-knockout mice, GATA-2 down-regulation may play a role in the reduction of a stem cell pool observed in AA. On the other hand, GATA-2 has been shown to be essential for the maintenance of immaturity of preadipocytes. If a pathological immune response in AA decreases the level of GATA-2 expression in not only hematopoietic stem cells but also stromal preadipocytes, it may accelerate the maturation of preadipocytes, leading to the formation of fatty bone marrow. To explore this possibility, the phenotypic change of stromal preadipocytes by suppression of GATA-2 was examined in this study. (Method) The GATA-2 expression level was suppressed by using siRNA for GATA-2 in mouse stromal preadipocyte cell lines, TBR9 and TBR343. After the treatment with siRNA, the adipocyte differentiation was induced by the incubation with insulin and dexamethasone for 7days. Then, the maturation level was examined by oil drops formation judged by oil red staining, and by the expression level of adipcin and PPAR-γ mRNA. Supporting activity of hematopoietic colony formation was also evaluated by using mouse fetal liver cells after siRNA treatment. (Results) By using designed siRNA, the GATA-2 expression was suppressed to 30% of control, whereas the expression level of GATA-3, which is co-expressed in preadipocytes, was unchanged. When GATA-2 was suppressed by siRNA, the oil drop formation and adipocyte-specific gene expression was significantly accelerated in both of stromal cells. Furthermore, the number of fetal liver hematopoetic colonies was significantly decreased by suppression of GATA-2, suggesting that GATA-2 down-regulation in stromal preadipocytes results in not only the acceleration of the maturation but also the reduced supporting activity of hematopoietic colony formation (Conclusion) These results suggest that suppression of GATA-2 in hematopoietic tissues induces the characteristic features of AA, i.e., decreased the number of hematopoietic stem cells and increased number of mature adipocytes.

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Human Hematopoietic Stem Cells and Leukemic Cells Form Cadherin-Catenin Based Junctional Complexes with Mesenchymal Stromal Cells

Blood ◽

10.1182/blood.v112.11.1367.1367 ◽

2008 ◽

Vol 112 (11) ◽

pp. 1367-1367 ◽

Cited By ~ 1

Author(s):

Patrick Wuchter ◽

Rainer Saffrich ◽

Wolfgang Wagner ◽

Frederik Wein ◽

Mario Stephan Schubert ◽

...

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Hematopoietic Stem Cells ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Leukemia Cells ◽

Feeder Layer ◽

Hematopoietic Stem ◽

Knock Down ◽

Junctional Complexes

Abstract The interaction between human hematopoietic stem cells (HSC) and their niche plays a key role in regulating maintenance of “stemness” and differentiation. We have demonstrated that a feeder layer of human mesenchymal stromal cells (MSC) can serve as a surrogate model for the niche for human HSC. We could also show, MSC are intimately connected to one another by a novel kind of adhering junction, consisting of villiformto-vermiform cell projections (processus adhaerentes). With this background, we have analyzed the intercellular junctional complexes between HSC and MSC. In comparison, we also studied the cell-cell contacts between leukemia cells (LC) and MSC. MSC were derived from bone marrow aspirates from healthy voluntary donors. HSC were isolated from umbilical cord blood. Leukemia cells that were CD34+ were obtained from bone marrow aspirates from patients suffering from acute myeloid leukemia at the time point of initial diagnosis. After 24–48 hours of co-cultivation, we stained the cellular contacts with a panel of antibodies specific for various components of tight, gap and adherens junctions. Using advanced confocal laser scanning microscopy in combination with deconvolution and volume rendering software, we were able to produce 3D-images of intercellular junctions between HSC/MSC as well as between LC/MSC. To examine the specific function of N-cadherin, we analyzed the effect of siRNA knock down of N-cadherin in MSC upon co-cultures of HSC and MSC. Intercellular connections between HSC and MSC are mainly characterized by podia formation of the HSC linking to the adjacent MSC. At the intimate contact zone to the MSC, we have identified the cytoplasmic plaque proteins alpha- and beta-catenin, co-localized with the transmembrane glycoprotein N-cadherin. Additionally, we compared these findings with a similar setting consisting of human LC co-cultured with feeder-layer of MSC. Our results demonstrated that in comparison to HSC, the proportion of leukemia cells adherent to the feeder-layer is significantly lower and podia formation is less frequent (ratio 1:3). However, the mechanism of adhesion through cadherin-catenin-complex has remained the same. At a functional level, we found that siRNA knock down of N-cadherin in MSC resulted in decreased adhesion of HSC to MSC and in a reduction of cell divisions of HSC. These results confirm that direct cellular contact via N-cadherin-based junctions is essential for homing and adhesion of HSC to the cellular niche and subsequently for the regulation of self-renewal versus differentiation in HSC.

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Kinetics of hematopoietic stem cells in the bone marrow of hepatectomized mice

Bulletin of Experimental Biology and Medicine ◽

10.1007/bf00800133 ◽

1978 ◽

Vol 85 (2) ◽

pp. 221-222

Author(s):

T. V. Osipova ◽

V. M. Bukhman ◽

G. Ya. Svet-Moldavskii

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Hematopoietic Stem Cells ◽

Hematopoietic Stem ◽

Kinetics Of

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Simian Immunodeficiency Virus Infection of Hematopoietic Stem Cells and Bone Marrow Stromal Cells

JAIDS Journal of Acquired Immune Deficiency Syndromes ◽

10.1097/00126334-200405010-00002 ◽

2004 ◽

Vol 36 (1) ◽

pp. 553-561 ◽

Cited By ~ 7

Author(s):

Chang I. Lee ◽

Morton J. Cowan ◽

Donald B. Kohn ◽

Alice F. Tarantal

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Virus Infection ◽

Hematopoietic Stem Cells ◽

Simian Immunodeficiency Virus ◽

Stromal Cells ◽

Bone Marrow Stromal Cells ◽

Marrow Stromal Cells ◽

Hematopoietic Stem ◽

Immunodeficiency Virus

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Mechanisms Mediating the Inhibitory Effect of All-Trans Retinoic Acid on Primitive Hematopoietic Stem Cells in Human Long-Term Bone Marrow Culture

Stem Cells ◽

10.1634/stemcells.18-3-214 ◽

2000 ◽

Vol 18 (3) ◽

pp. 214-219 ◽

Cited By ~ 10

Author(s):

J. Sammons ◽

N. Ahmed ◽

M. A. Khokher ◽

H. T. Hassan

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Retinoic Acid ◽

Hematopoietic Stem Cells ◽

Inhibitory Effect ◽

Bone Marrow Culture ◽

Hematopoietic Stem ◽

Trans Retinoic Acid ◽

All Trans Retinoic Acid

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Murine serum obtained from bone marrow-transplanted mice promotes the proliferation of hematopoietic stem cells by co-culture with MS-5 murine stromal cells

Growth Factors ◽

10.1080/08977190500361762 ◽

2006 ◽

Vol 24 (1) ◽

pp. 55-65 ◽

Cited By ~ 4

Author(s):

Akemi Nakayama ◽

Haruna Matsui ◽

Teruaki Fukushima ◽

Hiroshi Ichikawa ◽

Kensuke Yamada ◽

...

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Hematopoietic Stem Cells ◽

Stromal Cells ◽

Hematopoietic Stem

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Native associations of early hematopoietic stem cells and stromal cells isolated in bone marrow cell aggregates

Blood ◽

10.1182/blood.v83.2.361.bloodjournal832361 ◽

1994 ◽

Vol 83 (2) ◽

pp. 361-369 ◽

Cited By ~ 1

Author(s):

PE Funk ◽

PW Kincade ◽

PL Witte

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Stem Cell ◽

Hematopoietic Stem Cells ◽

Stromal Cells ◽

Stromal Cell ◽

Hematopoietic Cells ◽

Hematopoietic Stem ◽

Factor C

In suspensions of murine bone marrow, many stromal cells are tightly entwined with hematopoietic cells. These cellular aggregations appear to exist normally within the marrow. Previous studies showed that lymphocytes and stem cells adhered to stromal cells via vascular cell adhesion molecule 1 (VCAM1). Injection of anti-VCAM1 antibody into mice disrupts the aggregates, showing the importance of VCAM1 in the adhesion between stromal cells and hematopoietic cells in vivo. Early hematopoietic stem cells were shown to be enriched in aggregates by using a limiting-dilution culture assay. Myeloid progenitors responsive to WEHI-3CM in combination with stem cell factor (c-kit ligand) and B220- B-cell progenitors responsive to insulin-like growth factor-1 in combination with interleukin-7 are not enriched. We propose a scheme of stromal cell-hematopoietic cell interactions based on the cell types selectively retained within the aggregates. The existence of these aggregates as native elements of bone marrow organization presents a novel means to study in vivo stem cell-stromal cell interaction.

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