LFA-1-Mediated Adhesion Of Human Hematopoietic Progenitors To Bone Marrow Stromal Cells Enhances B-Lineage Differentiation Toward CD19+ ProB Cells, While The Generation Of CD7+CD56-CD45RA+ Multipotent Lymphoid and CD10+CD19- B-Biased Progenitors Is Supported By Soluble Factors Produced From Stromal Cells

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 3697-3697
Author(s):  
Hirohito Minami ◽  
Kohshi Ohishi ◽  
Yoshiki Nakamori ◽  
Masahiro Masuya ◽  
Naoyuki Katayama
Keyword(s):  
Bone Marrow ◽  
Stromal Cells ◽  
Bone Marrow Stromal Cells ◽  
Culture Conditions ◽  
Marrow Stromal Cells ◽  
Cell Contact ◽  
Hematopoietic Progenitors ◽  
Monocytic Cells ◽  
Lineage Differentiation ◽  
B Lineage

Abstract The regulatory mechanism of human lymphoid differentiation remains less defined. Here we examined how bone marrow stromal cells regulate human early lymphoid differentiation, using human telomerized bone marrow stromal cells that support the generation of CD7+CD56- early T and CD10+CD19+ proB cells from human hematopoietic progenitors. To examine the role of direct contact between hematopoietic progenitors and stromal cells in lymphopoiesis, cultures were performed by inhibiting the cell-cell contact with microporous insert or by incubating hematopoietic progenitors with conditioned medium collected from stromal cell cultures. The separation suppressed B-lineage differentiation to CD10+CD19+ cells, while the generation of CD7+ cells was not significantly influenced. The CD7+ cells generated with or without direct contact with stromal cells similarly had multipotent differentiation capacity for T, B, NK, granulocytic, and monocytic cells but not for erythroid cells in various culture conditions. On the other hand, even CD10+CD19- immature cells had more limited differentiation capacity for T, B, and monocytic cells in various culture conditions, and mostly differentiated toward CD10+CD19+ proB cells on the stromal cells. By time course analysis after coculture on the stromal cells, CD7+CD10- followed by CD10+CD19- and then CD10+CD19+ cells were developed. Some portion of CD7+CD10- and most of CD7-CD10+CD19- cells, upon recultured on stromal cells, differentiated toward CD10+CD19+ cells, but such B-lineage differentiation on the stromal cells was diminished by reculture with conditioned medium. ICAM-1 was expressed on the telomerized stromal cells. Coculture on stromal cells in the presence of LFA-1 neutralizing antibody that blocks the binding to ICAM-1 inhibited the differentiation to CD19+ proB cells. Our findings show that stromal cells support the generation of CD7+ multipotent lymphoid and CD10+ B-biased progenitors by producing soluble factors, but enhances B-lineage differentiation toward CD19+ proB cells in part via LFA-1-mediated direct cell-cell contact. Disclosures: No relevant conflicts of interest to declare.

Dependency of CLL Cell Survival on Direct Physical Interaction with Bone Marrow Stromal Cells In Vitro Is Largely Outweighed by Secretion of Soluble Factors with Time In Culture

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 1365-1365 ◽  
Author(s):  
Iris Gehrke ◽  
Simon Jonas Poll-Wolbeck ◽  
Michael Hallek ◽  
Karl-Anton Kreuzer
Keyword(s):  
Bone Marrow ◽  
Cell Survival ◽  
Stromal Cells ◽  
Bone Marrow Stromal Cells ◽  
Culture Conditions ◽  
Marrow Stromal Cells ◽  
Physical Contact ◽  
Physical Interaction ◽  
Soluble Factors

Abstract Abstract 1365 The major pathophysiology feature of chronic lymphocytic leukemia (CLL) cells is their extended life span due to a pronounced resistance towards apoptotic stimuli in vivo. Despite this, CLL cells die within a few days when isolated from their natural microenvironment and are placed under cell culture conditions. That is why the bone marrow microenvironment has been ascribed an essential role in maintenance of the apoptotic resistance of the CLL cell. Thereby both, the physical interaction between bone marrow stromal cells and CLL cells and the secretion of soluble factors have been described to be essentially involved. We analysed the survival capacity of CLL cells in monoculture and in coculture with the bone marrow-derived stromal cell line HS5 with and without physical separation using transwells for up to 7 days by flow cytometric determination of Annexin-V/PI status. As expected, in vitro CLL cell survival was significantly reduced when physical contact between CLL cells and bone marrow stromal cells was prevented. Interestingly, this was only the case for short term cultivation for up to three days. With time under culture conditions CLL cell survival became less dependent on direct physical contact with the HS5 feederlayer, suggesting the secretion of soluble factors to compensate for the loss of pro-survival signals obtained from direct cell-cell interactions over time. This was further supported by the fact of reduced survival support for CLL cells when HS5 proliferation, hence production and secretion of soluble factors, was prevented by mitomycin treatment or formaldehyde fixation. The use of an expanded human Cytokine Antibody Array (Affymetrix), which analyses the presence of the most common 36 cytokine proteins, might offer information about the composition of soluble factors present in the supernatant which are essential for CLL cell survival in vitro. In conclusion, while direct cell-cell contact between CLL cells and bone marrow stromal cells provides an immediate protection against in vitro apoptosis of CLL cells, the secretion of soluble factors, most likely by both, CLL and bone marrow stormal cells, leads to the creation of an in vitro environment which can to a certain extent compensate for the loss of prosurvival signals obtained by direct physical interactions. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Impaired Ability of Bone Marrow Stromal Cells to Support B-Lymphopoiesis With Age

Blood ◽  
1998 ◽  
Vol 91 (1) ◽  
pp. 75-88 ◽  
Author(s):  
Robert P. Stephan ◽  
Colette R. Reilly ◽  
Pamela L. Witte
Keyword(s):  
Bone Marrow ◽  
Stromal Cells ◽  
Bone Marrow Stromal Cells ◽  
Lymphoid Cells ◽  
Marrow Stromal Cells ◽  
Cell Contact ◽  
B Lymphopoiesis ◽  
Interleukin 7 ◽  
Age Related ◽  
B Lymphoid

B-lymphopoiesis decreases with age. We studied how aging affects bone marrow stromal cells, because they provide the growth factors and cell contacts required for B-lymphopoiesis. No differences were noted in the cell-surface phenotype of young and old primary-cultured stromal cells. Fluorescence-activated cell sorter-purified stromal cells from old mice were deficient in the ability to support the proliferation of interleukin-7 (IL-7)–specific B-lymphoid cell lines. The kinetics of this response indicated that IL-7 was not immediately available from stromal cells of either age and was further delayed on aged stromal cells. The levels of IL-7 protein within stromal cells were equivalent between young and old animals, suggesting that the production of IL-7 was not altered by aging. Negligible amounts of IL-7 were found either freely secreted or in the extracellular matrix of cultures of young and old marrow. Contact between the lymphoid cells and the primary stromal cells was required for detectable proliferation, suggesting that cell contact was required for the release of IL-7. We propose that stromal cells regulate B-lymphopoiesis by limiting the amount of IL-7 available to the developing precursors. Therefore, we conclude that the age-related decrease in the function of bone marrow stromal cells is related to the impaired release of IL-7.

scholarly journals Gfi1 expressed in bone marrow stromal cells is a novel osteoblast suppressor in patients with multiple myeloma bone disease

Blood ◽  
2011 ◽  
Vol 118 (26) ◽  
pp. 6871-6880 ◽  
Author(s):  
Sonia D'Souza ◽  
Davide del Prete ◽  
Shunqian Jin ◽  
Quanhong Sun ◽  
Alissa J. Huston ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Bone Marrow ◽  
Bone Disease ◽  
Stromal Cells ◽  
Bone Marrow Stromal Cells ◽  
Trichostatin A ◽  
Transcriptional Repressor ◽  
Marrow Stromal Cells ◽  
Cell Contact ◽  
Differentiation Markers

Abstract Protracted inhibition of osteoblast (OB) differentiation characterizes multiple myeloma (MM) bone disease and persists even when patients are in long-term remission. However, the underlying pathophysiology for this prolonged OB suppression is unknown. Therefore, we developed a mouse MM model in which the bone marrow stromal cells (BMSCs) remained unresponsive to OB differentiation signals after removal of MM cells. We found that BMSCs from both MM-bearing mice and MM patients had increased levels of the transcriptional repressor Gfi1 compared with controls and that Gfi1 was a novel transcriptional repressor of the critical OB transcription factor Runx2. Trichostatin-A blocked the effects of Gfi1, suggesting that it induces epigenetic changes in the Runx2 promoter. MM-BMSC cell-cell contact was not required for MM cells to increase Gfi1 and repress Runx2 levels in MC-4 before OBs or naive primary BMSCs, and Gfi1 induction was blocked by anti–TNF-α and anti–IL-7 antibodies. Importantly, BMSCs isolated from Gfi1−/− mice were significantly resistant to MM-induced OB suppression. Strikingly, siRNA knockdown of Gfi1 in BMSCs from MM patients significantly restored expression of Runx2 and OB differentiation markers. Thus, Gfi1 may have an important role in prolonged MM-induced OB suppression and provide a new therapeutic target for MM bone disease.

CXCL12 secretion by bone marrow stromal cells is dependent on cell contact and mediated by connexin-43 and connexin-45 gap junctions

2011 ◽  
Vol 12 (5) ◽  
pp. 391-398 ◽  
Author(s):  
Amir Schajnovitz ◽  
Tomer Itkin ◽  
Gabriele D'Uva ◽  
Alexander Kalinkovich ◽  
Karin Golan ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Gap Junctions ◽  
Stromal Cells ◽  
Bone Marrow Stromal Cells ◽  
Connexin 43 ◽  
Marrow Stromal Cells ◽  
Cell Contact

scholarly journals Direct Contact with Stromal Cells in Association with SDF-1 Is Important for B-Lineage Differentiation Toward CD19+ ProB Cells from Human Hematopoietic Precursors, but Dispensable for Generation of CD7+CD45RA+ Multipotent Lymphoid Precursors

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 5138-5138
Author(s):  
Hirohito Minami ◽  
Kohshi Ohishi ◽  
Yoshiki Nakamori ◽  
Masahiro Masuya ◽  
Naoyuki Katayama
Keyword(s):  
Stromal Cells ◽  
Direct Contact ◽  
Conflicts Of Interest ◽  
Neutralizing Antibody ◽  
Hematopoietic Progenitors ◽  
Monocytic Cells ◽  
Differentiation Potential ◽  
Human Cd34 ◽  
Lineage Differentiation ◽  
B Lineage

Abstract The regulatory mechanism of human early B- and T- lymphoid differentiation has not been well studied. Coculture on telomerized human bone marrow stromal cells supported the generation of CD7+CD45RA+ multipotent precursors with differentiation potential for T-, B-, NK-lineage, and monocytic cells, CD10+CD19-CD45RA+ B-biased precursors, and CD10+CD19+CD45RA+ proB cells from human CD34+lin- hematopoietic progenitors. CD7+CD45RA+ and CD10+CD19- lymphoid precursors can be developed from hematopoietic progenitors by soluble factors produced from stromal cells, but the generation of CD19+ proB cells was reduced. Replating analysis showed that direct contact with stromal cells promoted B-lineage differentiation toward CD19+ proB cells from CD34+lin- cell-derived CD7+CD45RA+ and CD10+CD19- lymphoid precursors. SDF-1 is shown to be critical for B-lineage differentiation from studies of mice. SDF-1 was produced from the human telomerized stromal cells. Inhibition of binding to SDF-1 with neutralizing antibody against CXCR4 suppressed B-lineage differentiation to CD19+ proB cells from hematopoietic precursors, while the generation of CD7+CD45RA+ cells was not significantly affected. By replating analysis, anti-CXCR4 Ab inhibited the differentiation to CD19+ proB cells from CD7+CD45RA+ and CD10+CD19- lymphoid precursors on stromal cells. These data indicate that direct contact with stromal cells in association with SDF-1 produced from stromal cells is important for B-lineage differentiation toward CD19+ proB cells from CD7+CD45RA+ and CD10+CD19- lymphoid precursors but dispensable for differentiation toward CD7+CD45RA+ multipotent lymphoid precursors. Disclosures No relevant conflicts of interest to declare.

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