Matične ćelije opredeljene za granulocitno-monocitnu lozu kostne srži i periferne krvi svinja

The pig is widely used as a large animal model for biomedical research and could be an interesting experimental model for studies of the hematopoietic system and its response in physiological and pathological conditions. With the intention of using the pig as a large animal model in hematopoietic research, a clonal assay in methylcellulose was developed and standardized for detection of committed progenitors of the granulocyte-macrophage lineage from adult pig bone marrow and peripheral blood. Progenitor cells were stimulated to proliferate and differentiate in vitro by adding pig leukocyte conditioned medium (LCM) as a source of homologous growth factors. The number of CFU-GM (Colony Forming Unit -Granulocyte-Macrophage) directly depended on the concentration of LCM. The proliferative rate of CFU-GM progenitor cells was determined by the cytosine arabinoside suicide technique. The percentage of bone marrow and peripheral blood CFU-GM cells in S phase of the cell cycle was 34.7% and 22.2%, respectively. The data obtained regarding the number and characteristics of pig bone marrow and peripheral blood CFU-GM confirmed that the organization of the pig CFU-GM progenitor cell compartment is similar and comparable to that in miniature swine, other animal species and humans.Svinja je životinja koja se koristi kao model u različitim biomedicinskim istraživanjima, a mogia bi biti i interesantan model u ispitivanju fiziologije i patolofizioloških promena hematopoetskog sistema. U cilju razvoja eksperimentalnog modela svinje u istraživanju hematopoeze, stanadardizovan je esej za odredivanje i karakterizaciju opredeljenih matičnih ćelija granulocitno-monocitne loze iz kostne srži i krvi odrasle svinje. Stimulacija proliferacije i diferencijacije ovih matičnih ćelija postignuta je dodavanjem medijuma kondicioniranog leukocitima (LCM - Leukocyte conditioned medium) bogatog faktorima rasta. Broj CFU-GM (Colony forming unit- granulocyte-macrophage) je direktno zavisio od koncentracije LCM-a. Procenat CFU-GM ćelija u S fazi ćelijskog cikiusa odredjivan je tehnikom 'suicida' korišćenjem citozin arabinozida (Ara-C) i iznosio je 34.7% za CFU-GM iz kostne srži i 22.2% za CFU-GM iz periferne krvi. Podaci dobijeni za broj i karakteristike CFU-GM iz kostne srži i periferne krvi potvrđuju da je ovaj odeljak matičnih ćelija kod odraslih svinja organizovan na isti način kao i kod minijaturnih svinja, drugih vrsta životinja i ljudi

Efekti IL-17 na funkcionalnu aktivnost ćelija periferne krvi

Interleukin-17 (IL-17) is a proinflammatory cytokine produced mainly by activated CD4+ and CD8+ T cells, while its specific receptor is ubiquitously distributed. The inflammatory capacity of IL-17 is based on its ability to stimulate a wide range of stromal cells to produce and release a number of proinflammatory mediators, some with a known impact on hematopoiesis particularly granulopoiesis. Recent data indicate a role for IL-17 in the pathogenesis of several inflammatory diseases, transplant rejection and tumor growth. The purpose of this study was to determine functional responses including the respiratory burst, nitric oxide (NO) production, adhesiveness and metabolical activity/viability of human peripheral blood leukocytes (total white blood cells, mononuclear cells and granulocytes) from healthy donors in the presence of recombinant human (rh)IL-17. The obtained results showed that rhIL-17 did not induce significant changes in the respiratory burst, NO production, and metabolical activity of each peripheral blood cell fraction the tested, while a slight increase in phorbol-12-myristate-13-acetate (PMA) stimulated adhesiveness of granulocytes and mononuclear cells was noted. The absence of significant changes in tested functional activities of various peripheral blood cells suggests that IL-17 does not express its proinflammatory ability in steady-state, since the requirement for its action really does not exist.Interleukin 17 (IL-17) je proinflamatorni citokin koga produkuju aktivirane CD4+ i CD8+ T ćelije, dok je njegov receptor ubikvitarno distribuiran. Inflamatorni kapacitet IL-17 se zasniva na njegovoj sposobnosti da stimuliše širok spektar stromalnih ćelija da produkuju i oslobađaju različite proinflamatorne medijatore, među kojima neki imaju efekte na hematopoezu posebno granulopoezu. Dosadašnji podaci ukazuju na ulogu IL-17 u patogenezi različitih inflamatornih bolesti, odbacivanju transplanta i razvoju tumora. Cilj ovog rada je bio da se odrede funkcionalni odgovori, uključujući respiratorni prasak, produkciju azot monoksida (NO), adhezivnost i metaboličku aktivnost/vijabilnost različitih ćelija periferne krvi (ukupnih leukocita, mononuklearnih ćelija i granulocita) zdravih donora, u prisustvu IL-17. Dobijeni rezultati su ukazali da IL-17 ne dovodi do značajnih promena respiratornog praska, produkcije NO i metaboličke aktivnosti ćelija periferne krvi, ali da uzrokuje blago povećanje forbol-12-miristat-13-acetat (PMA) stimulisane adhezivnosti granulocita i mononuklearnih ćelija. Odsustvo značajnih promena u ispitivanim funkcionalnim aktivnostima različitih ćelija periferne krvi, ukazuje da IL-17 ne eksprimira proinflamatorno dejstvo kod zdravih osoba, jer najverovatnije i ne postoji potreba za njegovim delovanjem

Odnos između totalnog kapaciteta za vezivanje gvožđa i koncentracije transferina kod novorođene prasadi tretirane gvožđe-dekstranom

Serum iron concentration and iron saturation of transferrin (Trf) are measures of body iron stores after administration of iron supplements. In clinical and experimental research, the complex determination of Trf was replaced by the simple determination of total iron binding capacity (TIBC). The objective of this work was to define if TIBC could be an adequate measure for Trf in neonatal piglets after i.m. iron administration. Treated piglets received 150 mg of iron-dextran i.m. the first day of life, and were compared to the untreated control group. Prior to iron administration, as well as on days 2, 8 and 12 after iron administration, serum iron and TIBC concentration were analyzed by an automatized chemical analyzer and Trf was determined by densitometry of electrophoretic strips. Our results show that regardless of iron treatment, TIBC is not a measure of Trf concentration in neonatal piglets two days after birth. At day 8 of their life a high correlation coefficient of these two parameters was established in non-treated animals, while in iron-treated piglets the same correlation was established 12 days after iron treatment. Thus, we suggest that in neonatal piglets, TIBC could be used as a measure of Trf concentration only 12 days after i.m. iron treatment.Određivanje statusa gvožđa u organizmu jedinke posle primene određenog preparata ovog mikroelementa moguće je utvrditi određivanjem njegove koncentracije u serumu i zasićenja transferina (Trf) gvožđem. U kliničkoj i eksperimentalnoj praksi složeno određivanje koncentracije Trf zamenjeno je jednostavnim određ ivanjem ukupnog serumskog kapaciteta za vezivanje gvožđa (TIBC). Cilj ovog rada je bio da se na modelu porasta serumskog Fe po i.m. aplikaciji Fe-dextrana novorođenoj prasadi, utvrdi odnos TIBC i Trf po aplikaciji ovog mikroelementa, kako bi se utvrdilo da li visoke doze gvožđa u serumu utiču na vrednost TIBC kao mere za određivanje koncentracije Trf. Vrednosti za serumsko Fe, TIBC i Trf poređene su između grupe životinja koja je odmah po rođenju dobila 150 mg Fe-dextrana i kontrolne grupe u kojoj životoinje nisu tretirane Fe-dextranom. Krv je uzorkovana pre aplikacije Fe-dextrana, drugog, osmog i dvanaestog dana po aplikaciji preparata gvožđa. Koncentracija gvožđa u serumu i TIBC su određivani standarnim kliničkim biohemijskim analizama, dok je koncenracija Trf određena denzitometrijom elektroforetskih traka. Dobijeni rezultati ukazuju da bez obzira na primenu preparata Fe, TIBC nije adekvatna mera za Trf kod novorođene prasadi u prva tri dana po rođenju. Osmog dana života prasadi, utvrđen je visoki stepen korelacije ova dva parametra kod životinja koje nisu bile tretirane, dok je kod tretiranih jedinki taj stepen korelacije postignut dvanaestog dana. Na osnovu izloženih rezultata se može zaključiti da se kod novorođene prasadi TIBC može koristiti kao mera za Trf tek 12 dana nakon i.m. tretmana Fe-dextranom

Matične ćelije zubne pulpe i njihov potencijalni značaj u regenerativnoj medicini

To date, three types of dental stem cells have been isolated: Dental Pulp Stem Cells (DPSC), Stem Cells From Human Exfoliated Deciduous Teeth (SHED) and Immature Dental Pulp Stem Cells (IDPC). These dental stem cells are considered as mesenchymal stem cells. They reside within the perivascular niche of dental pulp. They are highly proliferative, clonogenic, multipotent and are similar to mesenchymal Bone Marrow Stem Cells (BMSC). Also, they have high plasticity and can be easy isolated. The expressions of the alkaline phosphatase gene, dentin matrix protein 1 and dentinsialophosphoprotein are verified in these cells. Analyses of gene expression patterns indicated several genes which encode extracellular matrix components, cell adhesion molecules, growth factors and transcription regulators, cell signaling, cell communication or cell metabolism. In both conditions, in vivo and in vitro, these cells have the ability to differentiate into odontoblasts, chondrocytes, osteoblasts, adipocytes, neurons, melanocytes, smooth and skeletal muscles and endothelial cells. In vivo, after implantation, they have shown potential to differentiate into dentin but also into tissues like bone, adipose or neural tissue. In general, DPSCs are considered to have antiinflammatory and immunomodulatory abilities. After being grafted into allogenic tissues these cells are ableto induce immunological tolerance. Immunosuppressive effect is shown through the ability to inhibit proliferation of T lymphocytes. Dental pulp stem cells open new perspectives in therapeutic use not only in dentin regeneration, periodontal tissues and skeletoarticular, tissues of craniofacial region but also in treatment of neurotrauma, autoimmune diseases, myocardial infarction, muscular dystrophy and connective tissue damages.Iz zubne pulpe su do danas izolovane tri populacije matičnih ćelija koje su označene kao matične ćelije zubne pulpe (engl. Dental Pulp Stem Cells, DPSC), matične ćelije iz eksfoliranih mlečnih zuba (engl. Stem Cells From Human Exfoliated Decidual Teeth, SHED) i nezrele matične ćelije zubne pulpe (engl. Immature Dental Pulp Stem Cells, IDPC). Sve matične ćelije zubne pulpe su ektomezenhimalnog porekla i lokalizovane su u perivaskularnoj niši. One se lako i efikasno izoluju, visoko su proliferativne, klonogene, multipotentne, ispoljavaju visok stepen plasticiteta i i slične su mezenhimalnim matičnim ćelijama koštane srži (BMSC). U njima je pokazana visoka ekspresija gena alkalne fosfataze, proteina 1 matriksa dentina i dentin-sijalofosfoproteina. Takođe, istaknuta je važnost u ovim ćelijama ekspresije više gena koji kodiraju sintezu komponenti ekstracelularnog matriksa, molekula ćelijske adhezije, faktora rasta, transkripcionih faktora, gena prenosa ćelijskih signala, ćelijske komunikacije i metabolizma. U uslovima in vitro ili in vivo ove ćelije mogu da se diferenciraju, s određenim međusobnim razlikama, u pravcu odontoblasta, hondrocita, osteoblasta, adipocita, neurona/glije, glatkih i skeletnih mišićnih ćelija, endotelnih ćelija i melanocita. U uslovima in vivo, nakon implantacije, pokazuju različit potencijal za formiranje dentina, ali i koštanog, masnog i nervnog tkiva. Generalno se smatra da DPSC imaju anti-inflamatorno dejstvo i ispoljavaju imunom-odulatorni efekat. Takođe, dovode do imunološke tolerancije ukoliko se implantiraju u alogena tkiva. Sposobnost inhibicije proliferacije T limfocita ukazuje na njihovo imunosupresivno dejstvo. Matične ćelije zubne pulpe otvorile su nove perspektive u terapijskoj primeni ovih ćelija ne samo u regeneraciji dentina, tkiva periodoncijuma i koštano-zglobnog tkiva kraniofacijalne regije, već i u lečenju neurotraume, autoimunskih oboljenja, infarkta miokarda, mišićne distrofije i oštećenja vezivnog tkiva

Kultivacija matičnih i progenitorskih ćelija hematopoeze iz kostne srži hrčka

Hamster, a hibernating animal, is an important experimental model in research on the influence of hypothermia on different physiological processes. A simple procedure for cultivation and identification of hamster hematopoetic stem cells (HSC) and hematopoetic progenitor cells (HPC) is a premise for a successful investigation upon hypothermia effects on hematopoiesis. The aim of this work was to evaluate the utilization of commercially available methylcellulose media (MC) and recombinant mouse and human cytokines for hamster HSC and HPC assays, in order to enable further studies on these cells. Hamster bone marrow mononuclear cells (BMMNC) were plated in MC containing cytokines that support mouse or human HPC growth. Also, BMMNC were resuspended in cytokine supplemented liquid media and incubated for 5 weeks with a four day monitoring of viable cell number. We demonstrated that hamster hematopoietic progenitor cells committed for erythroid lineage and myeloid lineage successfully formed recognizable colonies in both mouse and human MC, while multipotent progenitor cells formed colonies only in mouse MC. We also defined conditions for the evaluation of hamster HSC activity in liquid cultures, based on continuous 5 weeks HSC proliferation. The obtained results verify the utilization of mouse specific MC for further research on hamster HPC biology during hypothermia.Fiziološka hibernacija u koju hrčci ulaze prilikom izlaganja niskim temperaturama, čini ove životinje zanimljivim eksperimentalnim modelom za ispitivanje hematopoeze u uslovima hipotermije. Preduslov za ovo ispitivanje je postojanje jednostavne metode za kultivaciju i identifikaciju hematopoetskih ćelija hrčka. Cilj ovog rada je bio da se ispita mogućnost kultivacije progenitorskih ćelija hematopoeze hrčka u kompletnoj metil celulozi dizajniranoj za kultivaciju mišijih i humanih hematopoetskih ćelija, kao i da se odrede optimalni uslovi za kultivaciju matičnih ćelija hematopoeze hrčka u tečnoj kulturi. Mononuklearne ćelije kostne srži hrčka su posađene u metil celulozu i u tečnu kulturu. Oba medijuma su sadržala kombinacije rekombinantnih mišijih i/ili humanih citokina. Kolonije progenitorskih ćelija opredeljenih za mijelopoezu i opredeljenih za eritropoezu su se formirale u metil celulozi dizajniranoj za kultivaciju mišijih i humanih hematopoetskih ćelija, dok su se primitivnije kolonije sastavljene od oba tipa ćelija (mijeloidna i eritrocitna loza) formirale samo u metil celulozi dizajniranoj za kultivaciju mišijih hematopoetskih ćelija. Osim toga, populacija matičnih ćelija hematopoeze hrčka je proliferisala u tečnim kulturama tokom 5 nedelja bez znakova opadanja proliferativnog potencijala. Ova istraživanja pokazuju da se primenjene metode mogu uspešno koristiti za ispitivanje hematopoeze kod hrčka

Karakterizacija matičnih ćelija izolovanih iz zubne pulpe mlečnih zuba dece

Background/Aim. The last decade has been profoundly marked by persistent attempts to use ex vivo expanded and manipulated mesenchymal stem cells (MSCs), as a tool in different types of regenerative therapy. In the present study we described immunophenotype and the proliferative and differentiation potential of cells isolated from pulp remnants of exfoliated deciduous teeth in the final phase of root resorption. Methods. The initial adherent cell population from five donors was obtained by the outgrowth method. Colony forming unit-fibroblast (CFU-F) assay was performed in passage one. Cell expansion was performed until passage three and all tests were done until passage eight. Cells were labeled for early mesenchymal stem cells markers and analysis have been done using flow cytometry. The proliferative potential was assessed by cell counting in defined time points and population doubling time was calculated. Commercial media were used to induce osteoblastic, chondrogenic and adipogenic differentiation. Cytology and histology methods were used for analysis of differentiated cell morphology and extracellular matrix characteristics. Results. According to immunophenotype analyses all undifferentiated cells were positive for the mesenchymal stem cell markers: CD29 and CD73. Some cells expressed CD146 and CD106. The hematopoietic cell marker, CD34, was not detected. In passage one, incidence of CFU-F was 4.7 ± 0.5/100. Population doubling time did not change significantly during cell subcultivation and was in average 25 h. After induction of differentiation, the multicolony derived cell population had a tri-lineage differentiation potential, since mineralized matrix, cartilage-like tissue and adipocytes were successfully formed after three weeks of incubation. Conclusion. Altogether, these data suggest that remnants of deciduous teeth dental pulp contained cell populations with mesenchymal stem cell-like features, with a high proliferation and tri- lineage differentiation potential and that these cultures are suitable for further in vitro evaluation of cell based therapies.Uvod/Cilj. Prošla dekada je bila posebno obeležena naporima na polju korišćenja ex vivo razvijenih i usmeravanih mezenhimskih matičnih ćelija (MSCs), kao sredstva za različite tipove regenerativne terapije. Cilj ove studije bio je da se utvrdi imunofenotip i potencijal za proliferaciju i diferencijaciju ćelija izolovanih iz zubne pulpe mlečnih zuba dece eksfoliranih u periodu kada je koren zuba bio u poslednjoj fazi resorpcije. Metode. Primarna adherentna populacija ćelija poreklom od pet donora dobijena je metodom eksplanta. Prisustvo progenitorskih ćelija koje obrazuju kolonije fibroblasta (CFU-F) pokazano je u prvoj pasaži. Do treće pasaže ćelije su ekspandirane, a potom korišćene za analiziranje. Imunofenotip je određen korišćenjem protočne citometrije. Proliferativni potencijal i vreme udvajanja ćelija (PDT) u kulturi je definisano na osnovu apsolutnog broja ćelija na početku i na kraju svake pasaže. Posle tronedeljne kultivacije ćelija u komercijalnim medijumima za stimulaciju osteogeneze, hondrogeneze i adipogeneze, citološkim i histološkim metodama je određena morfologija ćelija i karakteristike vanćelijskog matriksa. Rezultati. Antigeni koji karakterišu mezenhimske matične ćelije CD29 i CD73 su bili eksprimirani na svim nediferenciranim ćelijama, dok su antigeni CD146 i CD106 bili eksprimirani na ograničenom broju ćelija. Antigen CD34 (karakterističan za ćelije hematopoetske loze) nije bio eksprimiran. Incidencija CFU-F bila je 4,7 ± 0,5/100 ćelija. PDT se nije menjao tokom osam pasaža i u proseku je iznosio 25 h. Posle tronedeljne stimulacije diferencijacije u kulturama sa adipogenim medijumom došlo je do stvaranja ćelija sa masnim kapljicama, a u kulturama sa osteogenim medijumom došlo je do formiranja vanćelijskog matriksa sa deponovanim kalcijumovim solima. U kulturama sa hondrogenim medijumom došlo je do stvaranja tkiva sličnog hrskavici i vanćelijskog matriksa sa glikozaminoglikanima i kolagenom II. Zaključak. Zubna pulpa mlečnih zuba dece sadrži ćelijsku populaciju koja odgovara mezenhimskim matičnim ćelijama prema svojim karakteristikama, ima visok proliferativni potencijal i potencijal da se diferencira u tri ćelijske linije što je čini pogodnom za dalje in vitro analize i evaluaciju ćelijske terapije

Uticaj intraperitonealne aplikacije toluena rastvorenog u propilen glikolu na eritropoezu pacova wistar soja

The potential toxic effect of toluene on erythropoiesis was investigated during the enhanced erythropoietic activity provoked by application of propylene glycol (known as a mild hemolytic agent) in adult female Wistar rats. The animals were treated daily for 3, 7 or 11 days, with an intraperitoneal dose of toluene dissolved in propylene glycol (T+PG) or propylene glycol (PG) alone. The effects of T+PG and PG on some hematological parameters in peripheral blood and bone marrow were evaluated at the time of sacrifice. The number of red blood cells (RBC), reticulocytes (Rt) hematocrit values (PCV), and hemoglobin concentration (Hb) were determined in peripheral blood samples using standard laboratory procedures. Total bone marrow nucleated cells (TBMNC) were counted and the myelogram was analyzed as well. The number of early bone marrow erythroid progenitor cells-Burst Forming Unit-Erythroid (BFU-E) was assessed using a colony forming assay on methylcellulose. Toluene dissolved in PG induced a decrease in RBC number PCV and Hb concentration and an increase within the bone marrow BFU-E and precursor erythroid cell compartments, as well as the percentage of peripheral blood Rt, indicating enhanced erythropoietic activity. Very similar changes occurred when PG was administered alone. Therefore, it can be assumed that short-term application of a low dose of toluene (3 mg/kg) dissolved in PG did not have a toxic effect during PG induced enhanced erythropoietic activity.U ovom radu su prikazani rezultati ispitivanja mogućeg toksičnog uticaja toluena na eritropoezu tokom pojačane eritropoetske aktivnosti, izazvane aplikacijom blagog hemolitičkog agensa propilen glikola, kod odraslih ženki pacova Wistar soja. Eksperimentalnim životinjama je jedanput dnevno, tokom 3 7 i 11 dana, intraperitonealno aplikovan toluen rastvoren u propilen glikolu (T+PG), odnosno samo propilen glikol (PG). Efekti T+PG i PG na hematološke parametre periferne krvi i kostne srži su određivani u vreme žrtvovanja životinja. Broj eritrocita, retikulocita, hematokritska vrednost i koncentracija hemoglobina su određivani u perifernoj krvi standardnim laboratorijskim procedurama. U kostnoj srži je određivan ukupan broj ćelija sa jedrom i ispitan odnos ćelija prekursora mijeloidne i eritroidne loze. Broj ranih opredeljenih matičnih ćelija za eritrocitopoezu (BFU-E) određen je in vitro, na osnovu broja kolonija formiranih iz ćelija kostne srži na polučvrstoj podlozi od metil celuloze. Toluen rastvoren u propilen glikolu je u perifernoj krvi izazvao pad broja eritrocita, pad hematokritske vrednosti smanjenje koncentracije hemoglobina, a doveo je do porasta udela retikulocita. U kostnoj srži je došlo do porasta broja BFU-E, kao i eritrocitnih prekursora, što ukazuje na pojačanu eritropoetsku aktivnost. Ispitivani parametri se nisu značajno razlikovali prilikom aplikacije samog propilen glikola u odnosu na vrednosti parametara dobijenih prilikom aplikacije toluena rastvorenog u propilen glikolu. Zbog toga možemo da pretpostavimo da kratkotrajna aplikacija niske doze toluena (3 mg/kg) nema toksičan efekat na eritropoezu tokom pojačane eritropoetske aktivnosti izazvane aplikacijom propilen glikola

Mikrosredina hematopoeze

Steady-state hematopoiesis takes place in the bone marrow permissive microenvironment, composed of stromal cells, as well as extracellular matrix (ECM) components and regulatory molecules, produced by both stromal and hematopoietic cells. Stromal cells are both mezenchymal (endothelial cells fibroblasts, adipocytes, osteoblasts, myocytes, adventitial reticular cells) and hematopoietic (macrophages) in origin, which together provide cell to cell interactions, matrix proteins and growth factors essential for the maintenance, growth and differentiation of hematopoietic stem and progenitor cells. The ECM components (collagen, laminin, fibronectin, thrombospondin proteoglycans, hemonectin and various proteinases participating in their remodeling) are involved in different biological functions such as cell adhesion, binding and presentation of various cytokines and regulation of cell growth. These components serve to localize hematopoietic cells in the specific bone marrow microenvironment and it is suggested that in combination with cytokines are crucial for their compartmentalization. The regulatory molecules of hematopoietic microenvironment are cytokines, which regulate the survival, proliferation and differentiation of hematopoietic cells and cell adhesion molecules, which are responsible for the localization of hematopoiesis to the bone marrow. The activity of cytokines may be greater when the cytokine is present in a membrane-bound or ECM-associated form. Hematopoietic cells could also regulate normal hematopoiesis in an autocrine/paracrine manner, since it was shown that these cells express and secrete various growth factors, cytokines and chemokines.Proces hematopoeze se odvija u definisanoj tkivnoj mikrosredini kostne srži koja ima ključnu ulogu u njegovoj regulaciji. Mikrosredina hematopoeze podrazumeva funkcionalno jedinstvo stromalnih ćelija i ćelijskih produkata (molekuli ekstracelularnog matriksa i regulatorni faktori), koji čine kompleksni molekularni milje u kome se ostvaruju specifične interakcije hematopoetskih ćelija i komponenti mikrosredine. Stromalne ćelije mezenhimalnog porekla (endotelne ćelije, fibroblasti, adipociti osteoblasti, miociti, adventicijske ćelije retikuluma) i nemezenhimalnog porekla (makrofage), deluju na matične ćelije hematopoeze direktno među ćelijskim interakcijama, kao i produkcijom i deponovanjem pojedinih komponenti kompleksnog ekstracelularnog matriksa, a takođe i produkcijom i koncentrovanjem lokalnih citokina i faktora rasta sa hematopoetskim efektima, obezbeđujući na taj način gotovo sve činioce neophodne za proliferaciju i diferencijaciju matičnih ćelija hematopoeze. Komponente ECM (kolagen, laminin, fibronektin, trombospondin, proteoglikani, hemonektin i drugi glikoproteini kao i proteolitički enzimi koji omogućavaju remodelovanje ECM) su uključene u ćelijsku adheziju, vezivanje i prezentaciju različitih citokina i regulaciju ćelijskog rasta. Smatra se da komponente ECM omogućavaju lokalizuju hematopoetskih ćelija u specifičnoj mikrosredini kostne srži, i da u kombinaciji sa citokinima imaju presudnu ulogu u formiranju specifičnih niša. Regulatorni faktori hematopoetske mikrosredine su citokini koji regulišu preživljavanje, proliferaciju i diferencijaciju hematopoetskih ćelija i ćelijski adhezivni molekuli koji su odgovorni za lokalizaciju hematopoeze u kostnoj srži i za posredovanje u fizičkoj vezi između hematopoetskih ćelija i stromalnog tkiva mikrosredine. Ovi molekuli se u mikrosredini kostne srži nalaze kao solubilni, ili vezani za membrane stromalnih ćelija ili za komponente ECM, što može da pojača njihovu aktivnost. Takođe i same hematopoetske ćelije učestviju u regulaciji procesa hematopoeze produkujući citokine koji ispoljavaju autokrino/parakrine efekte

Cytochemical & ultrastructural alteration of cytoplasmic granules of rat peripheral blood neutrophils induced by chronic alcoholism & malnutrition

The specific influence of malnutrition on the pathophysiologic changes induced by chronic alcoholism is controversial. In an attempt to determine and demarcate the effects of protein malnutrition from those produced by alcoholism and to evaluate the precise effect of alcohol per se on cytochemical and ultrastructural properties of rat polymorphonuclear neutrophil (PMN) granules, we investigated the influence of chronic protein malnutrition or chronic alcoholism alone and in combination, in rats, After a 4 month experimental period various PMN properties, such as cytochemical, morphometrical and ultrastructural, as well, as neutrophil functions were studied. It was found that the degree of damage of PMNs induced either by ethanol or protein malnutrition alone was similar whereas their combination led to worsening of all markers of PMN functional ability. Ultrastructural changes of neutrophil granules including reduction, redistribution and atypical accumulation as well as appearance of autophagic vacuoles, confirmed their alteration which was emphasised by the additive pathophysiological interaction of alcoholism and chronic hypoprotein malnutrition

Effect of IL-17 on in vitro hematopoietic progenitor cells growth and cytokine release in normal and post-irradiated murine bone marrow

The influence of recombinant human IL-17 on granulocyte-macrophage (CFU-GM) and erythroid (BFU-E and CFU-E) progenitors and the release of IL-1 alpha/beta, IL-6 and erythropoietin (EPO) was estimated in the bone marrow cells obtained from normal and sublethally irradiated mice. In normal mice IL-17 increased CFU-GM and BFU-E and reduced CFU-E derived colonies numbers and augmented release of IL-6 and EPO. In irradiated mice the effects of IL-17 on hematopoietic progenitors were lineage-dependent, as well as dependent on their stage of differentiation and the time after the irradiation. IL-17 had no major effects on CFU-GM on day 1 and 3, but decreased their number on day 2, while enhanced both BFU-E and CFU-E on day 1 and 2 after irradiation, whereas on day 3 its effect on erythroid progenitors was again as observed in normal mice. After irradiation, IL-17 increased the release of IL-1 alpha, IL-6 and EPO. The observed effects suggested the involvement of IL-17 in the regulation of hematopoiesis and indicated that its effects on both hematopoictic progenitors and cytokine release are dependent on the physiological/pathological status of the organism