Probing the Interaction Forces of Prostate Cancer Cells with Collagen I and Bone Marrow Derived Stem Cells on the Single Cell Level

Purified primitive progenitor/stem cells from bone marrow represent likely target populations for ex vivo expansion of stem cells to be used in high-dose chemotherapy or gene therapy. Whereas such primitive progenitor cells require combined stimulation by multiple cytokines for growth, some cytokines selectively promote viability rather than growth when acting individually. We investigated here for the first time the direct effects of cytokines on survival of primitive CD34+CD38− human bone marrow progenitor cells at the single-cell level. Interleukin-3 (IL-3) and the ligands for c-kit (KL) and flt3 (FL) had direct and selective viability-promoting effects on a small fraction of CD34+CD38− but not CD34+CD38+ progenitor cells. Interestingly, the recently cloned thrombopoietin (Tpo), although stimulating little growth, kept most CD34+CD38− progenitors viable after prolonged culture, maintaining twofold and fourfold more progenitors viable than KL and IL-3, respectively. A high fraction of these progenitors had a combined myeloid and erythroid differentiation potential, as well as capacity for prolonged production of progenitor cells under stroma-independent conditions. In addition, Tpo promoted viability of CD34+CD38− long-term culture-initiating cells, further supporting the idea that Tpo promotes viability of primitive human progenitor cells. Finally, Tpo suppressed apoptosis of CD34+CD38− cells in culture. Thus, the present studies show a novel effect of Tpo, implicating a potential role of this cytokine in maintaining quiescent primitive human progenitor cells viable.

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Dynamic real-time imaging of marrow-resident neutrophils interacting with human mesenchymal stem cells during systemic lipopolysaccharide challenge.

Journal of Clinical Oncology ◽

10.1200/jco.2013.31.15_suppl.e22158 ◽

2013 ◽

Vol 31 (15_suppl) ◽

pp. e22158-e22158

Author(s):

Alex Yee-Chen Huang ◽

Jay T. Myers ◽

Deborah Sim Barkauskas

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Single Cell ◽

Human Mesenchymal Stem Cells ◽

Whole Body ◽

Migration Behavior ◽

Single Cell Level ◽

Cell Level

e22158 Background: Human mesenchymal stem cells (hMSCs) have gained intense interest due to their immune-modulatory, tissue differentiating and homing properties to sites of inflammation and tumor microenvironment, as evidenced by more than 200 ongoing clinical trials using hMSCs in a variety of clinical settings. Despite evidence demonstrating the bio-distribution of infused hMSCs in target organs using static fluorescence imaging or whole-body imaging techniques, there is controversy regarding how hMSCs exert their biological effects, and very little is known about how they behave dynamically within host tissues on a single-cell level in vivo. Methods: We infused fluorescently labeled clinical-grade hMSCs into immune-competent mice in which neutrophils and monocytes express a second fluorescent marker under the Lysozyme M (LysM) promoter. The recipient mice were then subjected to serial 4-D (xyzt) imaging of the bone marrow cavity with intravital two-photon microscopy (TPM) during acute systemic Lipopolysaccharide (LPS) challenges to observe changes in MSC and neutrophil migration behavior. Results: We were able, for the first time, to capture dynamic interactions between and migration pattern of hMSCs and LysM+granulocytes in the bone marrow of live mice during systemic LPS challenge. Contrary to some published reports, many of the infused hMSCs remained intact despite repeated cellular contacts with host neutrophils. However, we also observed the destruction and subsequent phagocytosis of some hMSCs by surrounding granulocytes. Conclusions: Our imaging platform provides opportunities to gain insight into the biology and therapeutic mechanisms of hMSCs in vivo at a single-cell level within live hosts.

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Ability of Early Acting Cytokines to Directly Promote Survival and Suppress Apoptosis of Human Primitive CD34+CD38− Bone Marrow Cells With Multilineage Potential at the Single-Cell Level: Key Role of Thrombopoietin

Blood ◽

10.1182/blood.v90.6.2282 ◽

1997 ◽

Vol 90 (6) ◽

pp. 2282-2292 ◽

Cited By ~ 154

Author(s):

Ole J. Borge ◽

Veslemøy Ramsfjell ◽

Li Cui ◽

Sten E.W. Jacobsen

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Single Cell ◽

Progenitor Cells ◽

Bone Marrow Cells ◽

High Dose ◽

Single Cell Level ◽

Cell Level ◽

Differentiation Potential

Abstract Purified primitive progenitor/stem cells from bone marrow represent likely target populations for ex vivo expansion of stem cells to be used in high-dose chemotherapy or gene therapy. Whereas such primitive progenitor cells require combined stimulation by multiple cytokines for growth, some cytokines selectively promote viability rather than growth when acting individually. We investigated here for the first time the direct effects of cytokines on survival of primitive CD34+CD38− human bone marrow progenitor cells at the single-cell level. Interleukin-3 (IL-3) and the ligands for c-kit (KL) and flt3 (FL) had direct and selective viability-promoting effects on a small fraction of CD34+CD38− but not CD34+CD38+ progenitor cells. Interestingly, the recently cloned thrombopoietin (Tpo), although stimulating little growth, kept most CD34+CD38− progenitors viable after prolonged culture, maintaining twofold and fourfold more progenitors viable than KL and IL-3, respectively. A high fraction of these progenitors had a combined myeloid and erythroid differentiation potential, as well as capacity for prolonged production of progenitor cells under stroma-independent conditions. In addition, Tpo promoted viability of CD34+CD38− long-term culture-initiating cells, further supporting the idea that Tpo promotes viability of primitive human progenitor cells. Finally, Tpo suppressed apoptosis of CD34+CD38− cells in culture. Thus, the present studies show a novel effect of Tpo, implicating a potential role of this cytokine in maintaining quiescent primitive human progenitor cells viable.

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Inhibition of glypican-1 expression induces an activated fibroblast phenotype in a human bone marrow-derived stromal cell-line

Scientific Reports ◽

10.1038/s41598-021-88519-7 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Sukhneeraj P. Kaur ◽

Arti Verma ◽

Hee. K. Lee ◽

Lillie M. Barnett ◽

Payaningal R. Somanath ◽

...

Keyword(s):

Prostate Cancer ◽

Bone Marrow ◽

Cancer Cells ◽

Cancer Cell ◽

Stromal Cell ◽

Prostate Cancer Cell ◽

Bone Cells ◽

Human Bone ◽

Human Bone Marrow ◽

Prostate Cancer Cells

AbstractCancer-associated fibroblasts (CAFs) are the most abundant stromal cell type in the tumor microenvironment. CAFs orchestrate tumor-stromal interactions, and contribute to cancer cell growth, metastasis, extracellular matrix (ECM) remodeling, angiogenesis, immunomodulation, and chemoresistance. However, CAFs have not been successfully targeted for the treatment of cancer. The current study elucidates the significance of glypican-1 (GPC-1), a heparan sulfate proteoglycan, in regulating the activation of human bone marrow-derived stromal cells (BSCs) of fibroblast lineage (HS-5). GPC-1 inhibition changed HS-5 cellular and nuclear morphology, and increased cell migration and contractility. GPC-1 inhibition also increased pro-inflammatory signaling and CAF marker expression. GPC-1 induced an activated fibroblast phenotype when HS-5 cells were exposed to prostate cancer cell conditioned media (CCM). Further, treatment of human bone-derived prostate cancer cells (PC-3) with CCM from HS-5 cells exhibiting GPC-1 loss increased prostate cancer cell aggressiveness. Finally, GPC-1 was expressed in mouse tibia bone cells and present during bone loss induced by mouse prostate cancer cells in a murine prostate cancer bone model. These data demonstrate that GPC-1 partially regulates the intrinsic and extrinsic phenotype of human BSCs and transformation into activated fibroblasts, identify novel functions of GPC-1, and suggest that GPC-1 expression in BSCs exerts inhibitory paracrine effects on the prostate cancer cells. This supports the hypothesis that GPC-1 may be a novel pharmacological target for developing anti-CAF therapeutics to control cancer.

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Single-Cell Transcriptome Analysis as a Promising Tool to Study Pluripotent Stem Cell Reprogramming

International Journal of Molecular Sciences ◽

10.3390/ijms22115988 ◽

2021 ◽

Vol 22 (11) ◽

pp. 5988

Author(s):

Hyun Kyu Kim ◽

Tae Won Ha ◽

Man Ryul Lee

Keyword(s):

Stem Cells ◽

Single Cell ◽

Cell Fate ◽

Human Cell ◽

Transcriptome Analysis ◽

Single Cell Analysis ◽

Embryonic Stem ◽

Single Cell Level ◽

Cell Analysis ◽

Cell Level

Cells are the basic units of all organisms and are involved in all vital activities, such as proliferation, differentiation, senescence, and apoptosis. A human body consists of more than 30 trillion cells generated through repeated division and differentiation from a single-cell fertilized egg in a highly organized programmatic fashion. Since the recent formation of the Human Cell Atlas consortium, establishing the Human Cell Atlas at the single-cell level has been an ongoing activity with the goal of understanding the mechanisms underlying diseases and vital cellular activities at the level of the single cell. In particular, transcriptome analysis of embryonic stem cells at the single-cell level is of great importance, as these cells are responsible for determining cell fate. Here, we review single-cell analysis techniques that have been actively used in recent years, introduce the single-cell analysis studies currently in progress in pluripotent stem cells and reprogramming, and forecast future studies.

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