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PubMed1997

Cytokine-mediated expansion does not deplete cord blood cells with stem cell characteristics.

Cord blood (CB) has been successfully used to regenerate the hematopoietic system after myeloablative therapy. We investigated whether cytokine mediated expansion depletes CB of cells with stem cell characteristics. CB mononuclear cells (MNC) were enriched for quiescent (primitive) stem cells by incubation with 25 micrograms/ml 5-Fluorouracil (5-FU) and control CB MNC were incubated with media alone. Cells were then incubated for 7 days with Interleukin-1 (IL1)+IL3+Stem Cell Factor (SCF) and progenitor content, cell cycle status, nucleated cell count, immunophenotype and resistance to 25 micrograms/ml 5-FU (primitive stem cells) were evaluated before and after cytokine exposure. Incubation with IL1+IL3+SCF caused an increase (fold expansion) in committed (28.6 +/- 8.1), immature (5.8 +/- 1.8), and primitive progenitors (4.1 +/- 0.8) among control CB MNC compared to a decrease in committed progenitors (0 +/- 0) but an increase in both immature (8.4 +/- 4.8) and primitive progenitors (7 +/- 2.9) among 5-FU resistant CB MNC. An increase in the proportion of CD34+ cells occurred in both fractions. Expanded control CB MNC showed a significant increase in numbers of 5-FU resistant committed (p = 0.024), immature (p = 0.014) and primitive progenitors (p = 0.01) as compared with fresh CB MNC. Re-exposure of 5-FU resistant expanded CB MNC to 5-FU shows growth of some immature and primitive progenitors. Cytokine-mediated expansion of untreated and quiescent CB cells is possible and cytokine-mediated expansion does not deplete CB cells with stem cell characteristics.

Antigens, CD34↗
PubMedDate not supplied

Research and use of stem cells.

Stem cells have enormous potential in medicine. Stem cell research is allowedsometimes just for to a limited extent in a large minority of countries. Nevertheless, the research and use of stem cells should be encouraged.

Biomedical Research↗
PubMed1983

Epidermal stem cells.

Stem cells are by definition present in all self-renewing tissues and are believed to play a central role in cell growth and differentiation. Existing evidence suggests that a subpopulation of epidermal basal keratinocytes represents stem cells; however, these cells have never been positively identified. In this paper we review evidence that in monkey palm epidermis there exist two morphologically distinct subpopulations of basal keratinocytes that are spatially segregated. One population, located in the shallow rete ridges, is characterized by a cytoplasm filled with tonofilaments and a highly convoluted ("serrated") dermal-epidermal junction; these cells may play a role in anchoring the epidermis to the dermis. In contrast, the other population, located at the tips of deep rete ridges, is characterized by a "primitive" cytoplasm containing abundant melanosomes and a relatively flattened ("nonserrated") dermal-epidermal junction. Tritiated thymidine labeling experiments suggest that the nonserrated basal keratinocytes are slow-cycling; however, a highly proliferative population of keratinocytes can be identified immediately above these basal cells. These findings are consistent with the concept that the nonserrated basal keratinocytes may represent stem cells that give rise to suprabasally located, transient amplifying cells before undergoing terminal differentiation. Monkey palm epidermis provides a model system for further studies of primate epidermal stem cells.

Animals↗
PubMed2001

Stem cells, cancer, and cancer stem cells.

Stem cell biology has come of age. Unequivocal proof that stem cells exist in the haematopoietic system has given way to the prospective isolation of several tissue-specific stem and progenitor cells, the initial delineation of their properties and expressed genetic programmes, and the beginnings of their utility in regenerative medicine. Perhaps the most important and useful property of stem cells is that of self-renewal. Through this property, striking parallels can be found between stem cells and cancer cells: tumours may often originate from the transformation of normal stem cells, similar signalling pathways may regulate self-renewal in stem cells and cancer cells, and cancer cells may include 'cancer stem cells' - rare cells with indefinite potential for self-renewal that drive tumorigenesis.

Animals↗
PubMed2006

Molecular switches and developmental potential of adult stem cells.

Stem cell commitment and differentiation entails the successive loss of self-renewal and developmental potential, and results in the final restriction to a terminally differentiated mature cell type. Hematopoiesis, the development of blood cells from hematopoietic stem cells in bone marrow, is particularly well studied, and at different branching points within the hematopoietic system multiple developmental intermediates have been identified. Here we describe a Flt3+ CD11b+ multipotent progenitor that can be amplified in vitro by a specific cytokine combination to high cell numbers, and following adoptive transfer into syngeneic mice, it generates dendritic cells but also additional mature cell types. By employing gene expression profiling with DNA microarrays and knockout mouse models, we demonstrate that the helix-loop-helix (HLH) transcription factor Id2 (inhibitor of DNA binding/differentiation 2) acts as a molecular switch in development of Langerhans cells (LCs), the cutaneous contingent of dendritic cells (DCs), and of specific DC subsets and B cells.

Adult↗
PubMed2006

7. Transplantation of undifferentiated, bone marrow-derived stem cells.

Stem cell research has known an enormous development, and cellular transplantation holds great promise for regenerative medicine. However, some aspects, such as the mechanisms underlying stem cell plasticity (cell fusion vs true transdifferentiation) and the functional improvement after stem cell transplantation, are highly debated. Furthermore, the great variability in methodology used by several groups, sometimes leads to confusing, contradicting results. In this chapter, we review a number of studies in this area with an eye on possible technical and other difficulties in interpretation of the obtained results.

Animals↗
PubMed2006

Human muscle stem cells.

Stem cells are unspecialized cells that have been defined in many different ways but they have two important characteristics that distinguish them from other cells in the body. First, they can replenish their numbers for long periods through cell division. Second, after receiving certain chemical signals, they can produce, through asymmetric cell division, a progeny that can differentiate or transform into specialized cells with specific functions, such as heart, nerve or muscle. In recent years, stem cells have received much attention owing to their potential use in cell-based therapies for human neurodegenerative diseases such as Parkinson's disease, stroke and muscular dystrophies. However, many questions need to be resolved before stem cells with myogenic potential are used in clinical standard protocols.

Animals↗
PubMed2006

Surface functionalization of inorganic nano-crystals with fibronectin and E-cadherin chimera synergistically accelerates trans-gene delivery into embryonic stem cells.

Stem cells holding great promises in regenerative medicine have the potential to be differentiated to a specific cell type through genetic manipulation. However, conventional ways of gene transfer to such progenitor cells suffer from a number of disadvantages particularly involving safety and efficacy issues. Here, we report on the development of a bio-functionalized inorganic nano-carrier of DNA by embedding fibronectin and E-cadherin chimera on the carrier, leading to its high affinity interactions with embryonic stem cell surface and accelerated trans-gene delivery for subsequent expression. While only apatite nano-particles were very inefficient in transfecting embryonic stem cells, fibronectin-anchored particles and to a more significant extent, fibronectin and E-cadherin-Fc-associated particles dramatically enhanced trans-gene delivery with a value notably higher than that of commercially available lipofection system. The involvement of both cell surface integrin and E-cadherin in mediating intracellular localization of the hybrid carrier was verified by blocking integrin binding site with excess free fibronectin and up-regulating both integrin and E-cadherin through PKC activation. Thus, the new establishment of a bio-functional hybrid gene-carrier would promote and facilitate development of stem cell-based therapy in regenerative medicine.

Animals↗
PubMed1995

Selective potentiation of IGE-dependent histamine release from rat peritoneal mast cells by stem cell factor.

Effect of stem cell factor on histamine release from rat peritoneal mast cells was studied. Although stem cell factor did not evoke histamine release by itself, it clearly potentiated histamine release from sensitized mast cells caused by antigen, anti-IgE and concanavalin A. However, stem cell factor did not affect histamine release caused by compound 48/80, calcium ionophore A23187 and substance P. Although maximum potentiation of antigen-induced histamine release by stem cell factor was accomplished after 1-10 minute-preincubation, potentiation was decline after a longer incubation period. Potentiation of histamine release by phosphatidylserine and non-mast cells in the rat peritoneal cavity was incubation time-dependent. Potentiation by stem cell factor was additive to that by phosphatidylserine or non-mast cells. These results indicate that stem cell factor selectively potentiates IgE-dependent histamine release from rat peritoneal mast cells, and suggest that the mechanism involved is distinct from that of phosphatidylserine or non-mast cells in the rat peritoneal cavity.

Animals↗
PubMed2005

Expansive effects of aorta-gonad-mesonephros-derived stromal cells on hematopoietic stem cells from embryonic stem cells.

BACKGROUND: Hematopoietic stem cells (HSCs) give rise to all blood and immune cells and are used in clinical transplantation protocols to treat a wide variety of refractory diseases, but the amplification of HSCs has been difficult to achieve in vitro. In the present study, the expansive effects of aorta-gonad-mesonephros (AGM) region derived stromal cells on HSCs were explored, attempting to improve the efficiency of HSC transplantation in clinical practice. METHODS: The murine stromal cells were isolated from the AGM region of 12 days postcoitum (dpc) murine embryos and bone marrow (BM) of 6 weeks old mice, respectively. After identification with flow cytometry and immunocytochemistry, the stromal cells were co-cultured with ESCs-derived, cytokines-induced HSCs. The maintenance and expansion of ESCs-derived HSCs were evaluated by detecting the population of CD34+ and CD34+Sca-1+ cells with flow cytometry and the blast colony-forming cells (BL-CFCs), high proliferative potential colony-forming cells (HPP-CFCs) by using semi-solid medium colonial culture. Finally, the homing and hematopoietic reconstruction abilities of HSCs were evaluated using a murine model of HSC transplantation in vivo. RESULTS: AGM and BM-derived stromal cells were morphologically and phenotypically similar, and had the features of stromal cells. When co-cultured with AGM or BM stromal cells, more primitive progenitor cells (HPP-CFCs) could be detected in ESCs derived hematopoietic precursor cells, but BL-CFC's expansion could be detected only when co-cultured with AGM-derived stromal cells. The population of CD34+ hematopoietic stem/progenitor cells were expanded 3 times, but no significant expansion in the population of CD34+Sca-1+ cells was noted when co-cultured with BM stromal cells. While both CD34+ hematopoietic stem/progenitor cells and CD34+Sca-1+ cells were expanded 4 to 5 times respectively when co-cultured with AGM stromal cells. AGM region-derived stromal cells, like BM-derived stromal cells, could promote hematopoietic reconstruction and HSCs' homing to BM in vivo. CONCLUSIONS: AGM-derived stromal cells in comparison with the BM-derived stromal cells could not only support the expansion of HSCs but also maintain the self-renewal and multi-lineage differentiation more effectively. They are promising in HSC transplantation.

Animals↗
PubMed1993

Mouse skin is particularly susceptible to tumor initiation during early anagen of the hair cycle: possible involvement of hair follicle stem cells.

Stem cells are believed to be a necessary target of chemical carcinogens. Based on autoradiographic, ultrastructural, and biologic criteria, we have recently proposed that hair follicle stem cells reside not in the bulb, but in the upper outer root sheath in an area called the bulge. Proliferating cells have been shown to be more susceptible to tumor initiation, and we have recently demonstrated that cells in the bulge undergo transient proliferation during early anagen. Therefore, we theorized that mouse skin should be particularly susceptible to carcinogen application during early anagen phase. In this paper, we show that early anagen Swiss and Sencar mouse skin is indeed particularly susceptible to one- and two-stage chemical carcinogenesis, resulting in tumor yields one to five times those obtained with telogen-timed carcinogen application. Our findings implicate a possible involvement of the bulge cells as precursors to some of the skin cancers, and support the concept that these are stem cells. These observations also raise important questions about the cellular origins and biologic behavior of chemically induced murine skin tumors.

9,10-Dimethyl-1,2-benzanthracene↗
PubMed2003

[The generation of neural stem cells: induction of neural stem cells from embryonic stem (ES) cells].

Neural stem cells are considered the ultimate lineage precursors to all neurons and glia. Despite the significance of neural stem cells in the mammalian brain development, their ontogenesis remains unclear. We have established a colony-forming embryonic stem (ES) sphere assay, where ES cells were cultured in serum-free media in the presence of leukemia inhibitory factor (LIF) to form floating spheres. LIF-dependent ES cell-derived sphere cells showed self-renewal and neural multipotentiality, cardinal features of the neural stem cell, but retained some non-neural properties and broader potential. We dabbed the cells in the ES cell-derived sphere of primitive neural stem cells. LIF-dependent sphere-forming cells were also present in the epiblast of embryonic day 5.5-7.5 mouse embryos. The generation of the in vivo primitive neural stem cell was independent of Notch signaling but the activation of Notch pathway was necessary for the transition from the primitive neural stem cell to the neural stem cell. We propose that the neural stem cell originates from the pluripotent inner cell mass/epiblast cell via the primitive neural stem cell stage under the control of Notch signaling.

Animals↗
PubMed2004

[Therapeutic use of stem cells].

Stem cells display important capacities of self renewing, proliferation and differentiation. Because those present in the embryo have the more remarkable properties, their potential use in the therapy of until now incurable degenerative diseases have been envisioned. Embryonic stem (ES) cells are located in the inner mass of the balstocyst at early stages of the development. Even in long-term cultures they still retain their undifferentiated features. Under specific culture conditions, ES cells can be committed into a variety of differentiation pathways, giving rise to large amounts of cells corresponding to different tissues (neurones, cardiomyocytes, skeletal muscle, etc.). However, producing these tissues from already established ES cell lines would lead to immune rejection when transplanted to patients. To prevent this pitfall and using the expertise accumulated by animal cloning by nucleus transfer, it has been proposed to adapt this technique to human ES cells. The therapeutic cloning consists in transferring the nucleus of somatic stem cells isolated from the patient into an enucleated oocyte, to allow blastocyst development from which ES cells will be derived. From these stem cells, compatible tissues will be then produced. The problem is that it is in theoretically possible to reimplant the cloned blastocyst into a surrogate mother for obtaining a baby genetically identical to the donor. This is called reproductive cloning. This worrying risk raises important ethic and legal questions.

Cell Differentiation↗
PubMed2005

Adult stem cells and cancer stem cells: tie in or tear apart?

Stem cell research is one of the new frontiers of medical science. Because of the unique self-renewable ability and powerful potential to differentiate, stem cells can be viewed as the mother of all cells in the body and have been investigated as a possible tool for reversing the degeneration and damage on organs. Recently, successful isolating cancerous stem cells from leukemia, breast and brain cancers provide a new target for eliminate cancer; however, it hints an increasing caution in using adult stem cells for organ repair. Cancerous stem cells share the same properties of self-renewal and differentiation with normal stem cells, with the addition of similar phenotype of adult stem cells isolated from the same tissue. Some believe that cancerous stem cells are derived from mutation of the normal stem cells, whereas others suspect it to be from different origins. Further investigation of the intrinsic factor underlying the behavior of adult stem cells and cancerous stem cells will shed light on both the fields of tissue engineering and cancer therapy. In this review, recent progresses in the studies of adult stem cells and cancerous stem cells are summarized to facilitate a better understanding and elicit much attention in this field.

Animals↗
PubMed1999

Human and rat hepatic stellate cells produce stem cell factor: a possible mechanism for mast cell recruitment in liver fibrosis.

BACKGROUND/AIMS: Mast cell numbers are markedly increased in advanced liver fibrosis. Stem cell factor may recruit mast cells to the liver following injury as it induces mast cell proliferation, survival and differentiation from resident tissue precursors. This study examines stem cell factor production in human fibrotic liver and by hepatic stellate cells during culture in vitro. METHODS: Stem cell factor production was examined in human fibrotic livers by ELISA and in human and rat hepatic stellate cell cultures using reverse transcription-polymerase chain reaction (RT-PCR), Northern blotting, Western blotting and immunocytochemistry. Co-culture studies examined adhesion between hepatic stellate cells and purified mast cells. RESULTS: RT-PCR showed stem cell factor mRNA was more consistently expressed in fibrotic human livers relative to normal, and ELISA confirmed this by showing stem cell factor protein was significantly increased 2-3-fold in homogenates of human cirrhotic liver (primary biliary cirrhosis, primary sclerosing cholangitis) relative to normal. RT-PCR detected stem cell factor mRNA in human and rat hepatic stellate cells activated by culture on plastic. This was confirmed by Western blotting, which showed that freshly isolated hepatic stellate cells expressed relatively little 30 kD stem cell factor compared to late primary culture activated hepatic stellate cells (14 day) and passaged hepatic stellate cells. As assessed by fluorescence immunocytochemistry, stem cell factor protein was homogeneously expressed by populations of culture-activated rat hepatic stellate cells. During co-culture, purified human skin mast cells adhered to hepatic stellate cell monolayers on plastic, and this adherence was inhibited >50% by addition of antibodies against stem cell factor. CONCLUSIONS: Hepatic stellate cells activated in vitro produce stem cell factor. These cells may play an important role in recruiting mast cells to liver during injury and fibrosis.

Animals↗
PubMed2003

Hedgehog-GLI signaling regulates the behavior of cells with stem cell properties in the developing neocortex.

Stem cells are crucial for normal development and homeostasis, and their misbehavior may be related to the origin of cancer. Progress in these areas has been difficult because the mechanisms regulating stem cell lineages are not well understood. Here, we have investigated the role of the SHH-GLI pathway in the developing mouse neocortex. The results show that SHH signaling endogenously regulates the number of embryonic and postnatal mouse neocortical cells with stem cell properties, and controls precursor proliferation in a concentration-dependent manner in cooperation with EGF signaling. These findings identify a crucial mechanism for the regulation of the number of cells with stem cell properties that is unexpectedly conserved in different stem cell niches.

Animals↗
PubMed2006

Astrocytogenesis of embryonic stem-cell-derived neural stem cells: Default differentiation.

Neural stem cells differentiate from embryonic stem cells via formation of neural stem spheres under free-floating conditions in astrocyte-conditioned medium. Subsequent culture of neural stem spheres on an adhesive substrate with fibroblast growth factor-2 promotes the migration of neural stem cells onto the substrate, resulting in an increase in the number of cells. These embryonic stem cell-derived neural stem cells can be differentiated almost exclusively into astrocytes by withdrawing fibroblast growth factor-2 from the medium without any additional instructions.

Animals↗
PubMed2003

Inorganic mercury changes the fate of murine CNS stem cells.

Stem cells isolated from the central nervous system of both embryonic and adult mice can generate neurons and glia through the activation of different patterns of differentiation in dependence of exposure to appropriate epigenetic signals. On the other hand, environmental conditions might affect the proliferation, migration, and differentiation of these cells. We report here, for the first time, that inorganic mercury affects the proliferative and differentiative capacity of adult neuronal stem cells (ANSCs). Actually, inorganic mercury increases apoptosis in ASNC. Furthermore, in stem cell-derived astrocytes, high levels of the 70 kDa heat shock protein (HSP-70) occur, while the levels of GTP-beta-tubulin activity dramatically decrease. Interestingly, when induced to differentiate, inorganic mercury modifies morphological proprieties of astrocytes, while the neuron population is reduced. These results demonstrate that inorganic mercury produces toxicity in the ANSC-derived neuronal population and affects the biological properties of the glial-derived population.

Animals↗