Stem cell biology : a never ending quest for understanding

Stem cells (SC) research is an important part of biotechnology that could lead to the development of new therapeutic strategies. A lot of effort has been put to understand biology of the stem cells and to find genes and subsequently proteins that are responsible for their proliferation, self-renewal and differentiation. Different cytokines and growth factors has been used to expand stem cells, but no combination of these factors was identified that could effectively expand the most primitive stem cells. Recently, however, genes and receptors responsible for SC proliferation and differentiation have been described. Ligands for these receptors or these genes themselves are being already used for ex vivo expansion of stem cells and the first data are very promising. New markers, such as CXCR4 and CD133, have been discovered and shown to be present on surface of hematopoietic stem cells. The same markers were recently also found to be expressed on neuronal-, hepatic- or skeletal muscle-stem cells. By employing these markers several laboratories are trying to isolate stem cells for potential clinical use. New characteristics of stem cells such as transdifferentiation and cell fusion have been described. Our team has identified a population of tissue committed stem cells (TCSC). These cells are present in a bone marrow and in other tissues and they can differentiate into several cell types including cardiac, neural and liver cells

The role of Insulin-like Growth Factor 1, Receptor Activator for Nuclear Factor κB ligand — Osteoprotegerin system, Interleukin 6 and 1β in post-transplantation bone metabolic disease in childhood

Wstęp: Znanych jest kilka czynników mogących mieć wpływ na zaburzenia kostne obserwowane często po przeszczepieniu komórekkrwiotwórczych (PKK). Ich dokładna etiopatogeneza pozostaje jednak wciąż nie znana, szczególnie u pacjentów po PKK przebytym wdzieciństwie. Celem pracy była ocena surowiczych stężeń: IGF-1, RANKL, OPG, IL-6 i IL1β oraz ich korelacji z gęstością mineralną kości(BMD) u dzieci i młodzieży po PKK.Materiał i metody: Do badań włączono 35 pacjentów w średnim wieku 8,48 ± 5,18 lat po allogenicznych (N = 21) i autologicznych (N = 14)PKK. Próbki krwi pobierano przed PKK, w dniu transplantacji, 3 i 6 miesięcy po PKK, a następnie co rok przez 2–8 lat. Stężenia IGF-1,RANKL, OPG, IL-1β, IL-6 oznaczane były metodami immunochemicznymi. BMD całego ciała oceniane było metodą DEXA (dual-energyX-ray absorptiometry) 6 miesięcy po PKK, a następnie co rocznie.Wyniki: Stwierdzono ujemne wartości Z-core dla BMD u wszystkich pacjentów. Były one znacząco wyższe u pacjentów po autologicznychniż po allogenicznych PKK. Surowicze stężenia IGF-1 i IL-6 znamiennie zmieniały się w czasie po PKK. Stężenia IGF-1 wzrastały od 2.roku po PKK, zaś IL-6 narastały do 12. miesiąca po PKK. Obserwowano również dynamiczne, chociaż nieznamienne zmiany stężeń OPG iRANKL po PKK. Wykazano korelacje RANKL i IGF-1 z BMD. IL-6 korelowała pozytywnie z IL-1β, obie interleukiny nie korelowały z BMD.Wnioski: Stwierdzono istotne i dynamiczne zmiany czynników mogących mieć wpływ na metabolizm kostny po transplantacji.Sugeruje się, że surowicze stężenia RANKL i IGF-1 mogą być markerami metabolizmu kostnego po PKK u dzieci i młodzieży.Introduction: Bone disorders observed commonly after haematopoietic stem cells transplantation (HSCT) can be caused by several factors,but their detailed pathomechanism is still not well known, especially in childhood.The aim of this study was to evaluate: IGF-I, RANKL-OPG system, IL-6, and IL1β levels and their association with bone mineral density(BMD) in children and adolescents after HSCT.Material and methods: Thirty five patients after allogeneic (N = 21) and autologous (N = 14) HSCT, mean age 8.48 ± 5.18 years, wereincluded in the study. Blood samples were taken before HSCT, on the transplantation day, three and six months after HSCT, then eachyear after HSCT for 2–8 years. RANKL, OPG, and IL-1β, IGF-1, and IL-6 were measured by immunochemistry. Total BMD was evaluatedsix months after HSCT using dual energy X-ray absorptiometry, then annually.Results: All Z-core values for BMD were negative in all patients. It was significantly higher in patients after auto HSCT than after allo HSCT.Serum levels of IGF-1 and IL-6 significantly changed after HSCT. IGF-I levels started to increase in the second year after transplantation.IL-6 increased up to 12 months after transplantation. Dynamic although not significant changes of OPG and RANKL levels were observedafter HSCT. RANKL and IGF-1 values correlated with BMD. IL-6 correlated positively with IL-1β but both did not correlate with BMD.Conclusions: Our data indicates that factors influencing bone remodelling change dynamically in the post-transplantation period.It suggests that serum RANKL and IGF-1 levels could be markers of bone metabolism after HSCT in paediatric patients

Heterogeneous populations of bone marrow stem cells--are we spotting on the same cells from the different angles?

Accumulated evidence suggests that in addition to hematopoietic stem cells (HSC), bone marrow (BM) also harbors endothelial stem cells (ESC), mesenchymal stem cells (MSC), multipotential adult progenitor cells (MAPC), pluripotent stem cells (PCS) as well as tissue committed stem cells (TCSC) recently identified by us. In this review we discuss the similarities and differences between these cell populations. Furthermore, we will present the hypothesis that all of these versatile BM derived stem cells are in fact different subpopulations of TCSC. These cells accumulate in bone marrow during ontogenesis and being a mobile population of cells are released from BM into peripheral blood after tissue injury to regenerate damaged organs. Furthermore, since BM is a "hideout" for TCSC, their presence in preparations of bone marrow derived mononuclear cells should be considered before experimental evidence is interpreted simply as trans-differentiation or plasticity of HSC. Finally, our observation that the number of TCSC accumulate in the bone marrow of young animals and their numbers decrease during senescence provides a new insight into aging and may explain why the regeneration processes becomes less effective in older individuals

Heterogeneous populations of bone marrow stem cells : are we spotting on the same cells from the different angles?

Accumulated evidence suggests that in addition to hematopoietic stem cells (HSC), bone marrow (BM) also harbors endothelial stem cells (ESC), mesenchymal stem cells (MSC), multipotential adult progenitor cells (MAPC), pluripotent stem cells (PCS) as well as tissue committed stem cells (TCSC) recently identified by us. In this review we discuss the similarities and differences between these cell populations. Furthermore, we will present the hypothesis that all of these versatile BM derived stem cells are in fact different subpopulations of TCSC. These cells accumulate in bone marrow during ontogenesis and being a mobile population of cells are released from BM into peripheral blood after tissue injury to regenerate damaged organs. Furthermore, since BM is a "hideout" for TCSC, their presence in preparations of bone marrow derived mononuclear cells should be considered before experimental evidence is interpreted simply as trans-differentiation or plasticity of HSC. Finally, our observation that the number of TCSC accumulate in the bone marrow of young animals and their numbers decrease during senescence provides a new insight into aging and may explain why the regeneration processes becomes less effective in older individuals

Evidence that platelet-derived microvesicles may transfer platelet-specific immunoreactive antigens to the surface of endothelial cells and CD34^+ hematopoietic stem/progenitor cells : implication for the pathogenesis of immune thrombocytopenias

The pathogenesis and tissue damage that accompanies destruction of platelets in immune thrombocytopenias (IT) is still not understood very well and in addition to platelets, other cells (e.g. endothelial cells, CD34+ hematopoietic stem/progenitors) may also become affected. Based on our previous work that platelet antigens (e.g., CD41) may be transferred by platelet-derived microvesicles (PMV) to the surface of other cells, we asked if platelet derived-antigens, especially those that are involved in the formation of anti-platelet antibodies in IT (e.g., against antigen HPA 1 a) could be also transferred by similar mechanism. To address this issue normal human CD34+ cells, human umbilical vein-endothelial cells (HUVEC) and monocytic cell line THP-1 were incubated with PMV derived from HPA1a+ donors. We noticed that the HPA1a antigen is highly expressed on PMV-derived from the HPAla positive platelets and is transferred in PMV-dependent manner to the surface of CD34+ cells, HUVEC and monocytic THP-1 cells. These cells covered with HPA1a positive PMV but not by PMV derived from HPAla negative platelets reacted with anti-HPA1a antibodies derived from the alloimmunized pregnant women. More importantly, human hematopoietic cells that were preincubated with HPA1a+ PMV and subsequently exposed to anti-HPA 1 a serum and human NK cells, become subject to elimination by antibody dependent cell cytotoxicity ADCC. Thus, we postulate that PMV-dependent transfer of antigens may playing an important role in "expanding" the population of target cells that may be affected by anti-platelet antibodies and explain several pathologies that accompany IT (e.g. damage of endothelium, cytopenias)

Evidence that platelet-derived microvesicles may transfer platelet-specific immunoreactive antigens to the surface of endothelial cells and CD34+ hematopoietic stem/ progenitor cells--implication for the pathogenesis of immune thrombocytopenias.

The pathogenesis and tissue damage that accompanies destruction of platelets in immune thrombocytopenias (IT) is still not understood very well and in addition to platelets, other cells (e.g. endothelial cells, CD34+ hematopoietic stem/progenitors) may also become affected. Based on our previous work that platelet antigens (e.g., CD41) may be transferred by platelet-derived microvesicles (PMV) to the surface of other cells, we asked if platelet derived-antigens, especially those that are involved in the formation of anti-platelet antibodies in IT (e.g., against antigen HPA 1 a) could be also transferred by similar mechanism. To address this issue normal human CD34+ cells, human umbilical vein-endothelial cells (HUVEC) and monocytic cell line THP-1 were incubated with PMV derived from HPA1a+ donors. We noticed that the HPA1a antigen is highly expressed on PMV-derived from the HPAla positive platelets and is transferred in PMV-dependent manner to the surface of CD34+ cells, HUVEC and monocytic THP-1 cells. These cells covered with HPA1a positive PMV but not by PMV derived from HPAla negative platelets reacted with anti-HPA1a antibodies derived from the alloimmunized pregnant women. More importantly, human hematopoietic cells that were preincubated with HPA1a+ PMV and subsequently exposed to anti-HPA 1 a serum and human NK cells, become subject to elimination by antibody dependent cell cytotoxicity ADCC. Thus, we postulate that PMV-dependent transfer of antigens may playing an important role in "expanding" the population of target cells that may be affected by anti-platelet antibodies and explain several pathologies that accompany IT (e.g. damage of endothelium, cytopenias)

An experimental survey of the production of alpha decaying heavy elements in the reactions of 238^{238}U +232^{232}Th at 7.5-6.1 MeV/nucleon

The production of alpha particle decaying heavy nuclei in reactions of 7.5-6.1 MeV/nucleon $^{238}$U +$^{232}$Th has been explored using an in-beam detection array composed of YAP scintillators and gas ionization chamber-Si telescopes. Comparisons of alpha energies and half-lives for the observed products with those of the previously known isotopes and with theoretically predicted values indicate the observation of a number of previously unreported alpha emitters. Alpha particle decay energies reaching as high as 12 MeV are observed. Many of these are expected to be from decay of previously unseen relatively neutron rich products. While the contributions of isomeric states require further exploration and specific isotope identifications need to be made, the production of heavy isotopes with quite high atomic numbers is suggested by the data.Comment: 12 pages, 12 figure

SDF-1 alone and in co-operation with HGF regulates biology of human cervical carcinoma cells

Stromal Derived Factor-1 (SDF-1)-CXCR4 axis plays a pivotal role in biology and metastasis of several tumors. The aim of this study was to see if SDF-1 alone or in combination with Hepatocyte Growth Factor (HGF) affects biology of human cervical carcinoma (HCC) cells. We found that HCC cell lines investigated in our study highly express CXCR4 on their surface. CXCR4 was also expressed on tumor cells in tissue sections derived from cervical cancer patients. At the same time normal cervical epithelium was negative for CXCR4 expression what suggests a strong correlation between CXCR4 and malignant cell phenotype. Subsequently, we studied a potential role of the SDF-1-CXCR4 axis in HCC and noticed that SDF-1 (i) chemoattracted HCC cells, (ii) enhanced their scattering, (iii) stimulated nuclear localization of beta-catenins and upregulated their target gene cyclin D1 and (iv) at the molecular level induced calcium flux and activated RAS-MAPK, PI3-AKT and JAK-STAT pathways. SDF-1-mediated functions were additionally enhanced in the presence of HGF. Thus, our data show that the SDF-1-CXCR4 axis affects biology of HCC cells. Furthermore, we postulate that this axis might become a potential target to prevent progression of cervical cancer

SDF-1 alone and in co-operation with HGF regulates biology of human cervical carcinoma cells.

Stromal Derived Factor-1 (SDF-1)-CXCR4 axis plays a pivotal role in biology and metastasis of several tumors. The aim of this study was to see if SDF-1 alone or in combination with Hepatocyte Growth Factor (HGF) affects biology of human cervical carcinoma (HCC) cells. We found that HCC cell lines investigated in our study highly express CXCR4 on their surface. CXCR4 was also expressed on tumor cells in tissue sections derived from cervical cancer patients. At the same time normal cervical epithelium was negative for CXCR4 expression what suggests a strong correlation between CXCR4 and malignant cell phenotype. Subsequently, we studied a potential role of the SDF-1-CXCR4 axis in HCC and noticed that SDF-1 (i) chemoattracted HCC cells, (ii) enhanced their scattering, (iii) stimulated nuclear localization of beta-catenins and upregulated their target gene cyclin D1 and (iv) at the molecular level induced calcium flux and activated RAS-MAPK, PI3-AKT and JAK-STAT pathways. SDF-1-mediated functions were additionally enhanced in the presence of HGF. Thus, our data show that the SDF-1-CXCR4 axis affects biology of HCC cells. Furthermore, we postulate that this axis might become a potential target to prevent progression of cervical cancer