The Use of Hoechst Staining To Identify Hematopoietic and Other Adult Stem Cells.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 4242-4242
Author(s):  
Rei Ogawa ◽  
Juri Fujimura ◽  
Hidemitsu Sugihara ◽  
Hidenori Suzuki ◽  
Hiko Hyakusoku ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Adipose Tissue ◽  
Hematopoietic Stem Cells ◽  
Adult Stem Cells ◽  
Surface Antigens ◽  
Adipose Derived Stem Cells ◽  
Cell Surface Antigens ◽  
Hematopoietic Stem ◽  
Hoechst Staining

Abstract BACKGROUND: Hoechst staining has been used to identify hematopoietic stem cells, but it may also be useful in identifying other adult stem cells. Here we report our efforts to purify and characterize stem cells in the stromal vascular fraction of adipose tissue with Hoechst staining, also comparing our results with those of our studies using bone marrow. METHODS: Stromal vascular fractions (SVFs) of adipose tissue and whole bone marrow (BM) were harvested from C57BL/6N mice, as were stem cells. Then the cells were stained with Hoechst 33342 and analyzed by flow cytometry and the number of cells in the side population (SP) counted. Moreover, surface antigens of SP cells were analyzed by flow cytometry using antibodies against CD44, 45, 45R, Sca-1, and c-kit, respectively, for 30 min on ice. Finally, the morphologic characteristics of cells in the SP of both BM and SVF were observed using electron microscopy. RESULTS: The percentage of SP cells in BM was about 0.05 to 0.1% and that in the SVF was about 1.0 to 3.0%. The cell-surface antigens of BM expressed were CD44 (−), CD 45 (+), CD 45R (−), Sca-1 (+) and c-kit (+), while those of SVF were CD44 (−), CD 45 (−), CD 45R (+/−), Sca-1 (+/−) and c-kit (−). Upon electron microscopic observation, both BM and SVF cells in the SP were considered to be remarkably immature (immature cell organelles and a high N/C ratio). CONCLUSION: The rate and expression patterns of cell-surface antigens in SP cells derived from BM were consistent with the results of previous reports. However, the same characteristics in SP cells derived from SVFs were clearly different. At present it is not clear whether cells in the SP of SVFs are adipose-derived stem cells. Indications were that there are 10 to 60 times as many immature cells in adipose tissue as in bone marrow. Moreover, it is possible that the great majority of cells in the SP of SVFs are not hematopoietic stem cells but unique adipose-derived stem cells. Finally, our studies suggest that Hoechst staining may be useful for identifying not only hematopoietic stem cells but also other adult stem cells.

scholarly journals Expression of CD34 marker in the adipose tissue derived stem cells

2018 ◽  
Vol 5 (1) ◽  
Author(s):  
Phuc Van Pham ◽  
Ngoc Bich Vu ◽  
Van Hong Tran
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Flow Cytometry ◽  
Mesenchymal Stem Cells ◽  
Adipose Tissue ◽  
Hematopoietic Stem Cells ◽  
Adipose Derived Stem Cells ◽  
Differentiation Potential ◽  
Hematopoietic Stem ◽  
Cd34 Expression

Introduction: Adipose-derived stem cells (ADSCs) are considered as mesenchymal stem cells (MSCs). Indeed, they display all characteristics of MSCs that compliant with the minimal criteria of MSCs suggested by Domonici et al. (2006). However, some recent studies showed that ADSCs contain the subpopulation that was positive with CD34 marker – a marker of hematopoietic stem cells. This study aimed to analyze and determine the expression of CD34 marker in ten samples of ADSCs obtained from 10 donors. Methods: All ADSC samples were isolated and expanded according to the published previous protocols. They were confirmed as the MSCs with some markers and differentiation potential, excepting the CD34 expression. Then they were cultured and analyzed the expression of CD34 by flow cytometry at passage 3, 5, 7 and 9. Results: The results showed that expression of CD34 in ADSCs was different between donors and their passages that accounted from 1.21% to 23.38%. Conclusion: These results suggested that ADSCs are not ‘truly” MSCs like MSCs from bone marrow.

scholarly journals The Tao of Hematopoietic Stem Cells: Toward a Unified Theory of Tissue Regeneration

2002 ◽  
Vol 2 ◽  
pp. 983-995 ◽  
Author(s):  
Kevin D. Bunting ◽  
Robert G. Hawley
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Hematopoietic Stem Cells ◽  
Adult Stem Cells ◽  
Developmental Plasticity ◽  
Marrow Cell ◽  
Cell Types ◽  
Therapeutic Benefit ◽  
Hematopoietic Stem ◽  
The Individual

Hematopoietic stem cells (HSCs) are the best studied of the tissue-specific stem cells. By definition, HSCs have long been regarded as restricted to formation of blood cells of both the lymphoid and myeloid lineages. HSCs residing in the bone marrow microenvironment have self-renewal capacity and can repopulate the hematopoietic system of irradiated transplant recipients for the lifetime of the individual. Therefore, HSCs are extremely important targets for gene therapy applications aimed toward the treatment of inherited and acquired blood disorders. However, recent studies have suggested that a subpopulation of HSCs may have the ability to contribute to diverse cell types such as hepatocytes, myocytes, and neuronal cells, especially following induced tissue damage. Preclinical amelioration of liver disease and myocardial infarcts by HSC-enriched bone marrow cell populations raises the possibility that HSC transplants have the potential to provide therapeutic benefit for a wide variety of diseases. These surprising findings contradict the dogma that adult stem cells are developmentally restricted. Extrapolation of these findings to the clinic will be facilitated by prospective identification of the stem cells that possess this developmental plasticity. Furthermore, characterization of the signaling pathways and molecular determinants regulating the remarkable transdifferentiation capacity of these stem cells may provide insight into novel approaches for modulating frequency of differentiative potential.

The Impact of Age and Gender on Hematopoietic Stem Cells and Immune Contexture of the Bone Marrow Microenvironment

2020 ◽  
pp. 1-6
Author(s):  
Rebar N. Mohammed
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Hematopoietic Stem Cells ◽  
Immune Cells ◽  
Absolute Number ◽  
Cell Number ◽  
Hematopoietic Stem ◽  
The Absolute ◽  
And Gender ◽  
The Impact

Hematopoietic stem cells (HSCs) are a rare population of cells that reside mainly in the bone marrow and are capable of generating and fulfilling the entire hematopoietic system upon differentiation. Thirty-six healthy donors, attending the HSCT center to donate their bone marrow, were categorized according to their age into child (0–12 years), adolescence (13–18 years), and adult (19–59 years) groups, and gender into male and female groups. Then, the absolute number of HSCs and mature immune cells in their harvested bone marrow was investigated. Here, we report that the absolute cell number can vary considerably based on the age of the healthy donor, and the number of both HSCs and immune cells declines with advancing age. The gender of the donor (male or female) did not have any impact on the number of the HSCs and immune cells in the bone marrow. In conclusion, since the number of HSCs plays a pivotal role in the clinical outcome of allogeneic HSC transplantations, identifying a younger donor regardless the gender is critical.

scholarly journals Influence of height on endothelial maintenance activity: a narrative review

2021 ◽  
Vol 26 (1) ◽  
Author(s):  
Yuji Shimizu ◽  
Takahiro Maeda
Keyword(s):  
Risk Factors ◽  
Cardiovascular Disease ◽  
Stem Cells ◽  
Bone Marrow ◽  
Hematopoietic Stem Cells ◽  
Inverse Association ◽  
Hematopoietic Stem ◽  
Genetic Characteristics ◽  
Maintenance Activity ◽  
Marrow Activity

AbstractRecent studies have revealed an inverse association between height and cardiovascular disease. However, the background mechanism of this association has not yet been clarified. Height has also been reported to be positively associated with cancer. Therefore, well-known cardiovascular risk factors, such as increased oxidative stress and chronic inflammation, are not the best explanations for this inverse association because these risk factors are also related to cancer. However, impaired blood flow is the main pathological problem in cardiovascular disease, while glowing feeding vessels (angiogenesis) are the main characteristic of cancer pathologies. Therefore, endothelial maintenance activity, especially for the productivity of hematopoietic stem cells such as CD34-positive cells, could be associated with the height of an individual because this cell contributes not only to the progression of atherosclerosis but also to the development of angiogenesis. In addition, recent studies have also revealed a close connection between bone marrow activity and endothelial maintenance; bone marrow-derived hematopoietic stem cells contribute towards endothelial maintenance. Since the absolute volume of bone marrow is positively associated with height, height could influence endothelial maintenance activity. Based on these hypotheses, we performed several studies. The aim of this review is not only to discuss the association between height and bone marrow activity, but also to describe the potential mechanism underlying endothelial maintenance. In addition, this review also aims to explain some of the reasons that implicate hypertension as a major risk factor for stroke among the Japanese population. The review also aims to clarify the anthropological reasons behind the high risk of atherosclerosis progression in Japanese individuals with acquired genetic characteristics.

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