The uterine environment in early pregnancy in the tammar wallaby

1996 ◽  
Vol 8 (4) ◽  
pp. 811 ◽  
Author(s):  
G Shaw
Keyword(s):  
Platelet Activating Factor ◽  
Fold Increase ◽  
Tammar Wallaby ◽  
Cell Contact ◽  
Embryonic Cells ◽  
Atp Production ◽  
Uterine Fluid ◽  
Uterine Environment ◽  
Uterine Secretions ◽  
Uterine Proteins

In tammar wallabies, Macropus eugenii, the uterine environment plays a key role in regulating development, because during the first two-thirds of gestation an acellular mucoid coat and shell prevent direct cell-cell contact between the endometrium and embryonic cells. This control is seen very clearly in the facultative lactational diapause of tammars. Removal of the suckled pouch young during the breeding season terminates diapause, leading to a distinct increase in metabolic activity of the embryo. By Day 4, oxidative metabolism of glucose has substantially increased, providing a four-fold increase in ATP production. By Day 5, RNA synthesis has increased. These changes are dependent on progesterone-induced changes in uterine secretions. By Day 3, there is greater progesterone secretion by the corpus luteum and, by Day 4, uterine protein synthesis has increased. The nature of the uterine regulatory factor is still not known. There are changes in some uterine proteins, but no detectable change in ionic components of the uterine fluid. Only one defined potential regulator, platelet-activating factor, has been identified, the concentration of which increased during reactivation. The influence of the steroid hormones progesterone and oestradiol on the uterus and diapausing embryo, and other changes that occur later in development, are also discussed in the present review.

scholarly journals Paf receptor expression in the marsupial embryo and endometrium during embryonic diapause

Reproduction ◽  
2014 ◽  
Vol 147 (1) ◽  
pp. 21-31 ◽  
Author(s):  
Jane C Fenelon ◽  
Geoff Shaw ◽  
Chris O'Neill ◽  
Stephen Frankenberg ◽  
Marilyn B Renfree
Keyword(s):  
Platelet Activating Factor ◽  
Tammar Wallaby ◽  
Receptor Expression ◽  
Limiting Factor ◽  
Embryonic Diapause ◽  
Paf Receptor ◽  
Sequential Activation ◽  
Uterine Environment ◽  
Low Levels

The control of reactivation from embryonic diapause in the tammar wallaby (Macropus eugenii) involves sequential activation of the corpus luteum, secretion of progesterone that stimulates endometrial secretion and subsequent changes in the uterine environment that activate the embryo. However, the precise signals between the endometrium and the blastocyst are currently unknown. In eutherians, both the phospholipid Paf and its receptor, platelet-activating factor receptor (PTAFR), are present in the embryo and the endometrium. In the tammar, endometrial Paf releasein vitroincreases around the time of the early progesterone pulse that occurs around the time of reactivation, but whether Paf can reactivate the blastocyst is unknown. We cloned and characterised the expression of PTAFR in the tammar embryo and endometrium at entry into embryonic diapause, during its maintenance and after reactivation. Tammar PTAFR sequence and protein were highly conserved with mammalian orthologues. In the endometrium, PTAFR was expressed at a constant level in the glandular epithelium across all stages and in the luminal epithelium during both diapause and reactivation. Thus, the presence of the receptor appears not to be a limiting factor for Paf actions in the endometrium. However, the low levels of PTAFR in the embryo during diapause, together with its up-regulation and subsequent internalisation at reactivation, supports earlier results suggesting that endometrial Paf could be involved in reactivation of the tammar blastocyst from embryonic diapause.

Biochemical studies of intrauterine components of the tammar wallaby Macropus eugenii during pregnancy

Development ◽  
1981 ◽  
Vol 62 (1) ◽  
pp. 325-338
Author(s):  
Elizabeth J. Thornber ◽  
Marilyn B. Renfree ◽  
Gregory I. Wallace
Keyword(s):  
Protein Concentration ◽  
Specific Binding ◽  
Polyacrylamide Gel Electrophoresis ◽  
Fold Increase ◽  
Tammar Wallaby ◽  
Macropus Eugenii ◽  
Tenfold Increase ◽  
Specific Proteins ◽  
Wet Weight

The in vitro uptake and incorporation of [3H]ui idine by blastocysts of the tammar wallaby showed a 16- and 30-fold increase from day 0 to day 10 after removal of pouch young, respectively. Two of the six non-expanded blastocysts recovered on day 5 showed a tenfold increase in incorporation. During the first ten days after removal of pouch young the diameter of the blastocyst increased threefold. Endometrial exudate from gravid uteri had a higher protein concentration than exudate from nongravid uteri (39·5 ± 0·9 and 32·0 ± 2·0 mg/ml (mean ± s.e.m.), respectively). Endometrial exudates from uteri where the blastocyst was actively growing were found to contain six uterine-specific proteins. These were separated by gradient polyacrylamide gel electrophoresis. Two of the proteins were pre-albumins and the others were larger molecules (M.W. 153000–670000). Two proteins were only present at particular stages of pregnancy: the other four were present at all stages from diapause to birth, in exudate from gravid and nongravid uteri. The specific binding of progesterone and androstenedione to proteins in endometrial exudates or uterine flushings from pregnant wallabies was less than one per cent of the value obtained from day-5 pregnant rabbits. The ability of mouse blastocysts to take up and incorporate [3H]uridine into acidinsoluble material increased threefold in the presence of day-10 endometrial exudates from wallabies. However, this was less than ten percent of the values obtained in the presence of bovine serum albumin. The concentration of calcium in endometrial exudates increased from 23·6 to 45·2 μg/ml during pregnancy; in endometrium it remained at 88·7 μg/g (wet weight) throughout pregnancy, and in plasma it was 53·3 μg/ml. The concentration of zinc in endometrial exudates was 4·5 μg/ml; in endometrium it decreased from 21·8 to 13·3 μg/g (wet weight) during pregnancy and in plasma it was 0·6 μg/ml.

Anoxia and adenosine induce increased cerebral blood flow rate in crucian carp

1994 ◽  
Vol 267 (2) ◽  
pp. R590-R595 ◽  
Author(s):  
G. E. Nilsson ◽  
P. Hylland ◽  
C. O. Lofman
Keyword(s):  
Blood Flow ◽  
Flow Rate ◽  
Crucian Carp ◽  
Blood Flow Rate ◽  
Fold Increase ◽  
Liver Glycogen ◽  
Glycolytic Flux ◽  
Brain Blood Flow ◽  
Atp Production ◽  
The Brain

The crucian carp (Carassius carassius) has the rare ability to survive prolonged anoxia, indicating an extraordinary capacity for glycolytic ATP production, especially in a highly energy-consuming organ like the brain. For the brain to be able to increase its glycolytic flux during anoxia and profit from the large liver glycogen store, an increased glucose delivery from the blood would be expected. Nevertheless, the effect of anoxia on brain blood flow in crucian carp has never been studied previously. We have used epireflection microscopy to directly observe and measure blood flow rate on the brain surface (optic lobes) during normoxia and anoxia in crucian carp. We have also examined the possibility that adenosine participates in the regulation of brain blood flow rate in crucian carp. The results showed a 2.16-fold increase in brain blood flow rate during anoxia. A similar increase was seen after topical application of adenosine during normoxia, while adenosine was without effect during anoxia. Moreover, superfusing the brain with the adenosine receptor blocker aminophylline inhibited the effect of anoxia on brain blood flow rate, clearly suggesting a mediatory role of adenosine in the anoxia-induced increase in brain blood flow rate.

Circus movements and blebbing locomotion in dissociated embryonic cells of an amphibian, Xenopus laevis

1976 ◽  
Vol 22 (3) ◽  
pp. 575-583
Author(s):  
K.E. Johnson
Keyword(s):  
Xenopus Laevis ◽  
Cell Contact ◽  
Embryonic Cells ◽  
Gastrula Stage ◽  
Isolated Cells ◽  
Daughter Cells ◽  
Cytoplasmic Protrusion ◽  
Endodermal Cells ◽  
In Cells

Circus movements, which involve the circumferential rotation of a hyaline cytoplasmic protrusion, occur in cells obtained by EDTA dissociation of gastrula-stage Xenopus laevis embryos. Only a few dissociated blastula-stage cells show circus movements, more early gastrula-stage cells show them, and nearly all late gastrula-stage cells show them. Circus movements cease in cells prior to mitosis and begin again in daughter cells after mitosis is completed. In early gastrulae, only 17% of prospective endodermal cells show circus movements while 79% of prospective mesodern, archenteric roof, and posterior neural ectoderm do so. Isolated cells as well as groups of cells in vitro are often propelled by circus movements. There is an obvious antagonism between cell contact and circus movements. The morphogenetic significance of circus movements and blebbing locomotion is discussed.

Abstract 585: Endothelial Cells from Adipose Tissue of Obese Humans Display Mesenchymal Characteristics

2016 ◽  
Vol 36 (suppl_1) ◽  
Author(s):  
Bronson A Haynes ◽  
Eric J Lehrer ◽  
Giann J Bhatt ◽  
Ryan W Huyck ◽  
Ashley N James ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Adipose Tissue ◽  
Prolonged Exposure ◽  
Endothelial Permeability ◽  
Fold Increase ◽  
Atp Production ◽  
Functional Changes ◽  
Twist 1

The mechanisms underlying vascular dysfunction in adipose tissue (AT) in obesity are not clearly understood. Our hypothesis is that in response to pro-inflammatory cytokines (PIC) present in obese AT, endothelial cells (EC) can de-differentiate and acquire a mesenchymal-like phenotype (EndoMT) that leads to endothelial dysfunction. To test our hypothesis, we measured endothelial and mesenchymal markers of CD31 + CD34 + EC isolated from omental (OM) and subcutaneous (SC) AT of bariatric subjects (BAMVEC) using RT-PCR and western blot. Permeability and oxidative metabolism were determined by ECIS and Seahorse analyzer XF e 24, respectively. BAMVEC isolated from both OM and SC fat showed very low protein expression of vWF and VE-Cadherin (EC markers) and abundantly expressed αSMA and the EMT transcription factor twist-1. To determine effects of PIC on EndoMT, commercially available primary endothelial cells from AT (HAMVEC) were treated in vitro with PIC (2.5ng/mL TNFα, IFNγ and TGFβ) for 1, 3 or 6 days. We found progressive down-regulation by >2-fold (p<0.001) of the EC markers vWF, VE-Cadherin, and Occludin compared to controls. As early as 1 day of PIC treatment twist-1 (p<0.001) and snail1 (p<0.05) showed an increase by >2-fold. Similarly, OM and SC BAMVEC expressed >2-fold increase in the mesenchymal genes twist-1, FSP1, αSMA, and snail1 compared to untreated HAMVEC. Metabolically, BAMVEC had increased ATP production and maximal respiration compared to HAMVEC suggesting increased oxidative phosphorylation, a marker of mesenchymal-like cells. PIC stimulation of HAMVEC yielded significant increases in endothelial permeability and motility (p<0.001). Notably, there were no significant differences in any of the markers between OM and SC BAMVEC. These results show that EC in obese AT exhibit a mesenchymal-like phenotype which may account for functional changes such as increased permeability and migration and are not depot specific. Using primary EC from human AT we showed that prolonged exposure to PIC induces a phenotype similar to CD31+CD34+ EC from obese AT. This supports the concept that AT inflammation can promote EC de-differentiation in vivo and our in vitro model is suitable for future studies to uncover the relevant mechanisms.

scholarly journals Regulation of Immunoglobulin G2 Production by Prostaglandin E2 and Platelet-Activating Factor

2000 ◽  
Vol 68 (3) ◽  
pp. 1563-1568 ◽  
Author(s):  
Yuichi Ishihara ◽  
Ji-Bo Zhang ◽  
Steve M. Quinn ◽  
Harvey A. Schenkein ◽  
Al M. Best ◽  
...  
Keyword(s):  
Prostaglandin E2 ◽  
Platelet Activating Factor ◽  
Pokeweed Mitogen ◽  
Cell Contact ◽  
Blood Leukocytes ◽  
Secreted Factors ◽  
Paf Antagonists ◽  
Localized Juvenile Periodontitis ◽  
Positive Regulators

ABSTRACT Patients with localized juvenile periodontitis (LJP) have elevated levels of immunoglobulin G2 (IgG2) in their sera. This is also observed in vitro when peripheral blood leukocytes from LJP patients are stimulated with pokeweed mitogen. In previous studies, we showed that lymphocytes from subjects with no periodontitis (NP subjects) produced substantial amounts of IgG2 when they were cultured with monocytes from LJP patients (LJP monocytes). These observations indicate that monocytes or monocyte-derived mediators are positive regulators of the production of IgG2. The present study was initiated to determine if secreted factors from LJP monocytes were capable of enhancing IgG2 production and to determine if prostaglandin E2 (PGE2), which LJP monocytes produce at elevated levels, enhances IgG2 production. Experiments in a transwell system and with monocyte-conditioned media indicated that cell-cell contact was not necessary for LJP monocytes to augment the production of IgG2 by T and B cells from NP subjects. Moreover, the production of IgG2 was selectively induced by the addition of PGE2 or platelet-activating factor (PAF), another lipid cytokine, which can elevate PGE2 synthesis. Furthermore, IgG2 production was abrogated when cells were treated with indomethacin, a cyclooxygenase inhibitor that blocks the synthesis of PGE2, or the PAF antagonists CV3988 and TEPC-15. The effects of indomethacin were completely reversed by PGE2, indicating that this is the only prostanoid that is essential for the production of IgG2. Similarly, PGE2 reversed the effects of a PAF antagonist, suggesting that the effects of PAF are mediated through the induction of PGE2 synthesis. Together, these data indicate that PGE2 and PAF are essential for the production of IgG2.

98 UTERINE INVOLUTION AND VASCULAR PERFUSION DURING EARLY POSTPARTUM IN MARES

2015 ◽  
Vol 27 (1) ◽  
pp. 142
Author(s):  
R. P. Arruda ◽  
K. M. Lemes ◽  
L. A. Silva ◽  
E. C. C. Celeghini ◽  
M. A. Alonso ◽  
...  
Keyword(s):  
Discrete Variables ◽  
Limited Evidence ◽  
Vascular Perfusion ◽  
Vascular Events ◽  
Fluid Content ◽  
Uterine Involution ◽  
Early Postpartum ◽  
Uterine Fluid ◽  
Uterine Environment ◽  
Main Effects

In horses, a rapid involution of the uterus occurs right after parturition, allowing the reestablishment of a favourable uterine environment for embryo development. However, limited evidence is found regarding the vascular events during puerperal period in mares. We aimed to evaluate the morphological (size of uterus and uterine fluid content) and haemodynamic (endometrial and mesometrial vascular perfusion) characteristics of the uterine involution process. Mares (n = 10) were evaluated by transrectal ultrasonography from the first day postpartum (d1) to the sixteenth day after first ovulation (D0 = ovulation). For ultrasound exams, a duplex B-mode and Doppler ultrasound instrument (M5 VET; Mindray Medical International Limited, China) equipped with a transrectal transducer was used. The previously pregnant (PH) and nonpregnant (NPH) horns were individually evaluated. Data were analysed for the main effects of horns (PH and NPH), day, and their interaction, using the PROC MIXED procedure of SAS software (9.3 version; SAS Institute Inc., Cary, NC, USA). Discrete variables were analysed by ANOVA. A reduction (P < 0.05) in the uterine diameter was observed during the first 7 days postpartum, but the rate of uterine involution (decrease in uterine size) decreased thereafter. The involution was complete around the d21 for the NPH and around d24 for the PH. Presence of uterine luminal fluid (mm) was increased between d1 (no fluid) and d2 (31.41 ± 2.88) postpartum, followed by a decrease between d4 (30.43 ± 4.52) and d7 (10.20 ± 1.76). No fluid was observed after d16 postpartum or after the third day postovulation (D3). For endometrial and mesometrial vascular perfusion, only a day effect (P < 0.05) was observed. An increase in the endometrial and mesometrial vascularization was detected, respectively, between d1 and d4, and between d1 and d2. Vascular perfusion did not differ after d4 for endometrial tissue, whereas it was reduced (P < 0.05) between d2 and d10 for mesometrium. For the vascular perfusion after ovulation, an increase (P < 0.05) from D0 to D5, followed by a decrease (P < 0.05) between D5 and D11 and an increase (P < 0.05) between D11 and D14 was observed in the endometrial and mesometrial tissues. The profile of the vascular perfusion in endometrium and mesometrium after first ovulation postpartum is similar to that observed during oestrous cycles and early pregnancy, indicating a return of the uterus to the pre-pregnant uterine characteristics in mares.Research was supported by FAPESP process no. 2010/10692-9 and CNPq process no. 135954/2011-8.

6 Decelerating embryo development? Characterisation of the uterine environment in European roe deer (Capreolus capreolus) during diapause

2019 ◽  
Vol 31 (1) ◽  
pp. 128
Author(s):  
V. A. van der Weijden ◽  
A. R. Vegas ◽  
V. Milojevic ◽  
A. B. Rüegg ◽  
J. T. Bick ◽  
...  
Keyword(s):  
Developmental Stage ◽  
Embryo Development ◽  
Developmental Stages ◽  
Roe Deer ◽  
Embryonic Diapause ◽  
Chromatography Tandem Mass Spectrometry ◽  
Uterine Fluid ◽  
Liquid Chromatography Tandem Mass ◽  
Uterine Environment ◽  
Differentially Abundant Proteins

The early developing embryo faces a continuously changing microenvironment to supports its growth. In the European roe deer, this environment accompanies embryonic diapause, a period of up to 4 months in which fertilization and subsequent implantation are decoupled. Diapause is characterised by a deceleration of embryonic growth. In most ruminants such as cattle and sheep, interferon tau (IFNt) plays a major role in maternal recognition of pregnancy. Uniquely to ruminants, the roe deer embryo does not secrete IFNt. The roe deer was used as a model species to gain insights into the changing uterine environment devoid of IFNt that supports prolonged decelerated embryo development, resumption of developmental velocity, and subsequent implantation. Uterine fluid samples from 188 female does were collected during regular huntings between September and January, and 4 developmental stages-blastocysts at early, mid, and late diapause and elongated embryos (16, 57, 97, and 18 does per developmental stage, respectively)-were defined. The developmental stages were assigned based on morphological characteristics of the embryo and the embryonic genomic DNA content. For the analysis of amino acids (AA), all 188 uterine fluid samples were subjected to targeted liquid chromatography-tandem mass spectrometry. Almost all AA increased over the course of embryo development. Although most AA showed developmental stage-specific concentration peaks, serine, glycine, alanine, glutamate, and glutamine were most abundantly present irrespective of the developmental progression. For the analysis of the protein abundances in the uterine fluid in a selected subset of samples (n=5 per developmental stage), holistic liquid chromatography-tandem mass spectrometry identified and quantified a total of 819 proteins with a false discovery rate of &lt;1%. Comparison between the developmental stages revealed 106 differentially abundant proteins. Most changes in protein abundance that occurred related to embryo elongation. Interestingly, 713 proteins remained stable during embryo development, indicating that these proteins may contribute to prolonged embryo survival during embryonic diapause. The differentially abundant proteins were clustered with DAVID Bioinformatics Resources 6.8 (https://david.ncifcrf.gov/). The most enriched clusters were cell-cell adhesion, biosynthesis of AA and carbon metabolism, microtubule, structural molecule activity, and chaperone binding. The ongoing detailed identification of stably abundant proteins will advance our basic understanding of the embryos’ needs for sustained survival during prolonged decelerated development. In addition, a comparison with the protein abundances around the time of maternal recognition of pregnancy in other species could advance our knowledge on conserved proteins that support embryo development and establishment of pregnancy in mammals. Our findings may contribute to defining optimal in vitro embryo culture conditions in a species-independent manner and potentially identify factors capable of halting embryo development.

Identification and regulation of the platelet-activating factor acetylhydrolase activity in the mouse uterus in early pregnancy

2001 ◽  
Vol 13 (6) ◽  
pp. 367 ◽  
Author(s):  
O. Chami ◽  
C. O'Neill
Keyword(s):  
Enzyme Activity ◽  
Early Pregnancy ◽  
Platelet Activating Factor ◽  
Density Lipoprotein ◽  
Mouse Uterus ◽  
Uterine Endometrium ◽  
Native Molecular Mass ◽  
Platelet Activating Factor Acetylhydrolase ◽  
Uterine Fluid ◽  
Detergent Treatment

Platelet-activating factor (PAF) is a product of the embryo and the endometrium in early pregnancy. The actions of PAF may be regulated by its degradation and this is largely achieved by the enzyme PAF acetylhydrolase (PAF:ah; EC 3.1.1.47). The present study characterized the PAF:ah in the endometrium and uterine fluid of mice during early pregnancy. The enzyme activity from uterine endometrium and luminal fluids had the same biochemical characteristics as the plasma form of the enzyme. The three sources of enzyme activity (i) had an apparent native molecular mass greater than 106 Da, but this was reduced after detergent treatment and purification to 60–65 kDa; (ii) bound to cholesterol hemisuccinate agarose matrix; and (iii) were found in the high density lipoprotein-enriched fraction after density gradient ultracentrifugation. In castrate females, oestradiol-17β (E2) caused a dose-dependent increase in the activity of the enzyme in endometrium and luminal fluid. Progesterone (P4) inhibited the E2-induced increase in PAF:ah in uterine tissue. Treatment with E2 alone caused an increase in endometrial PAF:ah activity within 24 h, which declined within 48 h. In luminal fluid, the same treatment caused increased activity within 24 h, peaking after 48 h of treatment and then declining. In E2-treated castrate females, mRNA for an intracellular (but not plasma) form of PAF:ah was detected, yet the intracellular form was not detected biochemically. The results suggest that most of the enzyme activity was not produced locally, but probably resulted from the influx of the plasma form of the enzyme.

Human Brain Endothelial Cells (HUBEC) Can Expand Both Human Bone Marrow and Cord Blood SCID-Repopulating Cells (SRC) through Cell Contact Rather Than Soluble Factors.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 36-36
Author(s):  
Naoko Takebe ◽  
Xiangfei Cheng ◽  
Ann M. Farese ◽  
Emily Welty ◽  
Barry Meisenberg ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Endothelial Cells ◽  
Human Brain ◽  
Ex Vivo ◽  
Fold Increase ◽  
Ex Vivo Expansion ◽  
Scid Mice ◽  
Cell Contact ◽  
Brain Endothelial Cells ◽  
Cd34 Cells

Abstract Human brain endothelial cells (HUBEC), a U.S. Navy proprietary cell line, was reported previously by Chute et al as a promising co-culture ex vivo expansion system for both adult bone marrow (ABM) and cord blood (CB) hematopoietic stem cells (HSC).a,b,c We report here our results of using HUBEC in ex vivo expansion and in vivo engraftment assay using NOD-SCID mice. CD34+ enriched fresh ABM was obtained using the method as described previously.a,b However, we used frozen CB and the same cytokines for both ABM and CB expansion whereas Chute et al used fresh CB and different cytokines. Ex vivo expansion studies for both ABM and CB were performed for 7 days in the HUBEC coated plates with previously reported cell density and cytokine cocktail containing GM-CSF, IL-3, IL-6, SCF, and flt-3 (GM36SF) in IMDM 10% FBS media.a HSC injections and BM harvesting of NOD-SCID mice as well as flow cytometric analysis were performed using the methods of Chute et al.a NOD-SCID mice were transplanted with limiting doses of either fresh ABM CD34+ cells or freshly thawed CB CD34+. The progeny of the identical doses of ABM CD34+ or the progeny of the identical doses of CB CD34+ cells was then transplanted. Culture with GM36SF alone resulted in a 15.5-fold and 70-fold increase in total cells, a 3.4-fold and 32-fold increase in CD34+ cells, and a 4.8-fold and 4.1-fold increase in CD34+/CD38- cells for ABM and CB, respectively. In contrast, HUBEC co-culture with GM36SF yielded a 25-fold and 48-fold increase in total cells, a 8.9-fold and 13-fold increase in CD34+ cells, and 114-fold and 106-fold increase in CD34+/CD38- cells for ABM and CB, respectively. HUBEC co-culture without GM36SF supported a 1.0-fold and 1.0-fold increase in total cells, a 0.06-fold and 0.1-fold increase in CD34+ cells, and 0.25-fold and 0.2-fold increase in CD34+/CD38- cells for ABM and CB. HUBEC co-culture with GM36SF and transwell (non-contact culture) resulted in a 20-fold and 48-fold increase in total cells, a 6-fold and 8-fold increase in CD34+ cells, and a 32-fold and 38-fold increase in CD34+/CD38- cells for ABM and CB. Overall, the transwell expansion of CD34+/CD38- population in both ABM and CB was reduced to 30% of that achieved in the contact culture. ABM CD34+ cells (5 x 105) engrafted 60% and the progeny of 5 x 105 cultured in the HUBEC monolayer with GM36SF engrafted in 90% of transplanted mice. CB CD34+ cells (1 x 104) engrafted 27% and the progeny 1 x 104 CB CD34+ cells cultured in the HUBEC monolayer with GM36SF engrafted 64% of NOD-SCID mice. SRC frequencies calculated as a 3.12-fold and 2.7-fold increase in CD34+ enriched ABM and CB, respectively, which was less than reported previously.a,b In summary, HUBEC supports and expands SRC mainly through cell-to-cell contact between HSC and endothelial cells, with HUBEC-secreted factors playing a minor role.

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