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1.
Hepatocytes derived from foetal rat liver synthesize and secrete albumin and transferrin when maintained in primary culture. These proteins are produced for at least seven days under the conditions of culture. Studies on hepatocyte cultures derived from 12, 13, 14, 15 and 19-day foetal rats show that the maximal cellular rate of secretion of both proteins increases about 50-fold over this period. The maximal rate of albumin secretion in all cultures is achieved after one day in culture and decreases in hepatocytes from early foetuses after the fourth to sixth day in culture. Transferrin secretion by hepatocytes from 12 to 15 day foetuses increases markedly during the second day of culture and is relatively constant thereafter. In contrast, secretion of transferrin by hepatocytes from 19-day foetuses is constant from the first day of culture. The results show that both albumin and transferrin are synthesized and secreted by the foetal liver as early as the twelfth day of gestation. The increase in the rate of transferrin secretion that occurs during culture of hepatocytes from 12 to 15 day foetuses may reflect the development of a secretory mechanism that is different from that for albumin. 相似文献
2.
The effect of dexamethasone on albumin production by fetal rat hepatocytes in culture 总被引:2,自引:0,他引:2
G C Yeoh V J Brighton D A Angus M Kraemer J Vassy M T Chalemeau 《European journal of cell biology》1985,38(1):157-164
Hepatocytes derived from 15 and 19-day gestation rats synthesize and secrete albumin during culture. Albumin secretion is maintained when the culture medium is supplemented with dexamethasone but declines in its absence. The fall in secretion rate correlates with the level of albumin messenger RNA in the respective cultures. Even when dexamethasone is present, the level of albumin production in 19-day gestation hepatocytes is 6 to 7 times greater than that observed in hepatocytes derived from 15-day gestation rats. Immunocytochemical studies were undertaken to establish whether the difference in secretion rate was due to a difference in the amount of albumin produced by all the hepatocytes of the respective cultures or whether there were fewer hepatocytes which were capable of synthesizing albumin in the less mature liver. The results indicate that albumin production is reduced in all hepatocytes when cultured in the absence of dexamethasone. 相似文献
3.
J I Raeside 《Biology of reproduction》1983,28(1):128-133
An influence of gonadotropins on steroid secretion by the early fetal ovary of the domestic pig was shown by organ culture and radioimmunoassay. Gonads from fetuses at Days 32-37 of gestation were cultivated singly for 9-12 days in biologically supplemented medium. One member of each pair of gonads was exposed to human chorionic gonadotropin (hCG) or ovine luteinizing hormone (LH), and the other served as a control. A marked stimulating effect on androgen secretion was noted with both gonadotropins. The major androgen found was androstenedione, with secretion rates of greater than 200 ng/gonad per 24 h for some explants exposed to hCG. Little or no androstenedione production occurred unless gonadotropin had been added to the culture medium. Lesser amounts of testosterone (usually less than 5% of the total of androstenedione and testosterone) were present. The data demonstrate a remarkable latent capacity for androgen biosynthesis by the early fetal pig ovary. 相似文献
4.
Ontogeny of proliferative and cytotoxic responses to interleukin 2 and concanavalin A in murine fetal thymus 总被引:1,自引:0,他引:1
The ontogeny of proliferative and cytotoxic responses to concanavalin A (Con A) and interleukin 2 (IL 2) in C57BL/6J (B6) fetal thymus (FT) was investigated. Embryonic thymocytes were either taken from embryos at different times of gestation or from 14 day B6 FT that were maintained as organ cultures for various times. It was found that the B6 FT could proliferate to Con A and EL4 SN (an IL 2 containing culture supernatant) in a synergistic fashion. This synergy between Con A and EL4 SN was first observed at the 16th to 17th day of gestation. A similar differentiation process took place in 14-day FT that had been maintained as organ cultures; the synergy between Con A and EL4 SN was first observed after 3 days in organ culture. This synergy increased with increasing time of organ culture, and was most evident after 10 days. The synergy between Con A and EL4 SN was also observed when the EL4 SN was replaced with IL 2 which had been purified from crude EL4 SN to apparent homogeneity. B6 FT could also form cytotoxic T lymphocytes (CTL) on stimulation with Con A and EL4 SN. Con A-activated CTL (polyspecific) were detected by including phytohemagglutinin in the assay medium. CTL response was first detected in the 17-day fetal thymus by using this assay. In organ cultures, CTL responses were first detected after 4 days in organ culture, and reached peak levels after 12 to 14 days. The CTL precursor (CTL-P) frequencies in the B6 FT after 2, 5, 10, and 14 days in organ culture were less than 1/10,000, 1/2232, 1/297, and 1/70, respectively; the corresponding CTL-P frequency in adult thymus was 1/60. After 6 days in organ culture, B6 FT could also form CTL in response to Con A and pure IL 2. This finding suggests that the ability to synthesize other differentiation factors that are required for CTL responses is acquired at an early time of thymic differentiation. 相似文献
5.
Ontogeny of the glucocorticoid receptor and its relationship to tyrosine aminotransferase induction in cultured foetal hepatocytes. 总被引:3,自引:3,他引:0
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The glucocorticoid receptor activity that can be detected in the liver from 15-day foetal rats would appear to be associated with the haemopoietic cells. In hepatocytes, purified by culture for 1-2 days from 15-day foetal rats, the glucocorticoid receptor activity is low and dexamethasone does not induce the enzyme tyrosine aminotransferase. If culture is continued both receptor activity and steroid responsiveness are acquired. Cultured hepatocytes from 19-day foetal liver contain receptor from the first day of culture and, furthermore, the subsequent level of response to glucocorticoids is directly correlated with the actual receptor concentration. It would appear that the glucocorticoid receptor is not acquired by hepatocytes until after 18 days of gestation. Nevertheless, the fact that bromodeoxyuridine has no effect on the rate of accumulation of receptor in hepatocytes suggests that the differentiative event leading to the subsequent appearance of the receptor has already occurred before day 15 of gestation. However, the acquisition of the receptor would appear to be dependent on mitosis as cytosine arabinoside can inhibit the process. 相似文献
6.
Kirsti Heinonen 《Biochimica et Biophysica Acta (BBA)/General Subjects》1975,399(1):113-123
At the 18th day of gestation and thereafter foetal rat liver explants in organ culture showed the competence to respond to dexamethasone by increased cystathionase activity, whereas the ability to respond to dibutyryl cyclic AMP or glucagon became evident at a later developmental stage (during the last 2 days prior to term). Simultaneous incubation with cycloheximide inhibited the stimulatory effect of these agents on foetal rat liver cystathionase activity in vitro. Dexamethasone and glucagon were both capable of increasing liver cystathionase activity both in newborn and 3-day-old animals in vivo. 相似文献
7.
G N Pluzhnikova N V Sadovnikova Ia Iu Kondrat'ev E I Lezhnev V P Lavrovskaia 《Biulleten' eksperimental'no? biologii i meditsiny》1986,102(7):86-88
The data are reported on albumin secretion by rat hepatocytes and insulin secretion by pancreatic beta-cells of newborn rats during cell cultivation on flat synthetic membrane in conditions of continuous medium perfusion. Albumin and insulin secretion by the appropriate cultures was higher in continuous medium perfusion than in the control. Enhanced sensitivity of pancreatic beta-cells to glucose, as compared to the control was revealed. It is concluded that continuous medium perfusion of hepatocytes and pancreatic beta-cells in the primary culture had a favourable effect on albumin and insulin secretion by the appropriate cultures. 相似文献
8.
Miroshnichenko IV Piatin VF Alekseeva AS Tiurin NL 《Rossi?skii fiziologicheski? zhurnal imeni I.M. Sechenova / Rossi?skaia akademiia nauk》2002,88(2):248-256
Power spectral analysis of inspiratory discharges of C3-C5 ventral roots in brainstem-spinal cord preparation from foetal (18 and 20 gestation days) and newborn (0-1 and 2-3 postnatal days) rats was performed. The respiratory centre perinatal development manifests itself by decreasing of respiratory rhythm variability and increasing of inspiratory burst duration. In foetal inspiratory bursts, low-frequency oscillations (1-10 Hz) dominate. In early postnatal stage, the relative power of low-frequency oscillations begin to decrease, and medium frequency oscillations (10-50 Hz) start to dominate over the inspiratory discharge. The data obtained suggests, that perinatal maturation of respiratory centre is characterised by stabilisation of the respiratory rhythm generation and developmental alteration of inspiratory activity's spectral and temporary parameters. 相似文献
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11.
The development of rat alpha 2-macroglobulin. Studies in vivo and in cultured fetal rat hepatocytes 总被引:1,自引:0,他引:1
S Fletcher T Thomas G Schreiber P C Heinrich G C Yeoh 《European journal of biochemistry》1988,171(3):703-709
During inflammation and tissue injury, there is an increase in the plasma concentration of several proteins, the acute-phase proteins. The levels of some acute-phase proteins have been reported to increase in pregnant and tumour-bearing animals. Rat alpha 2-macroglobulin is classified as an acute-phase protein. In this study we report the expression of alpha 2-macroglobulin in various tissues during development of the rat embryo by analysis of mRNA. The tissues studied are liver, visceral yolk sac, placental labyrinth, decidua and trophoblast. In addition, the sites of alpha 2-macroglobulin expression are localized by in situ hybridization of cDNA for alpha 2-macroglobulin to mid-sagittal cryosections of rat embryos. The level of mRNA coding for alpha 2-macroglobulin is determined in the liver of rats aged between 12 days gestation and 2 days postnatal. alpha 2-Macroglobulin mRNA is first observed in fetal liver from 12 days of gestation and increases after day 17, reaching a maximum on day 20. At this time the level is greater than that found in the liver of an adult rat suffering from acute inflammation. alpha 2-Macroglobulin mRNA is detectable in the yolk sac, placental labyrinth, trophoblast tissue and decidua. In the decidua the alpha 2-macroglobulin message is first detected at 8 days of gestation, with high levels observed from 10 to 21 days of gestation. These observations are supported by in situ hybridization studies. Experiments using cultured hepatocytes show that cells derived from rats at 15 days and 19 days of gestation are capable of synthesizing and secreting alpha 2-macroglobulin. Both synthesis and secretion can be induced by the addition of dexamethasone to the culture medium. 相似文献
12.
Liver cells of new-born rats, which were found to be able to form spheroidal aggregates when cultured on a nonadherent plastic substratum, were studied under various conditions of culture, mainly by adding different nutrients and growth factors to the culture medium. Analysis of hepatocyte-specific functions was carried out by immunoprecipitation to detect specific proteins newly secreted by liver cell spheroids on different days of culture. When no supplement was added to culture medium, the secretion of albumin and transferrin by liver cell spheroids was no longer detectable after 2 weeks of culture. When dexamethasone, glucagon, insulin, and EGF were added to culture medium, the secretion of albumin and transferrin remained detectable at least until 60 days of culture. This was even more striking when trace elements were added in addition to the three hormones and EGF. The effects of addition of these various factors to culture medium were also detectable with respect to alpha-FP secretion. Even after 54 days of culture in total supplemented medium, these liver cell spheroids could be transferred on a collagen-coated plastic substratum to form a monolayer of uniform liver parenchyma-like cells. The presence of extracellular matrix-like material was observed on the surface of cell spheroids. This could be responsible for attachment and fusion between cell spheroids. Thus, liver cell spheroids cultured in total supplemented medium ensured cell attachment to a biological matrix and cell-cell contact, which is thought to help maintain cell differentiation. Liver cell spheroids offer the possibility of toxicological and pharmacological studies as well as cultures in biomatrix and coculture systems. In addition these liver cells can be used for experiments in liver cell transplantation. 相似文献
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14.
Kinetic constants for liver glycogen synthase (UDPglucose: glycogen 4-alpha-D-glucosyltransferase, EC 2.4.1.11) with respect to UDPglucose have been measured in foetal liver homogenates from samples taken during late gestation (days 17-22) and the first hours after birth. The V of the inactive form of glycogen synthase increased markedly in this period and there was a significant increase in V of the active enzyme to a maximum at day 20 of gestation. The Km for UDPglucose measured in the presence of glucose-6-P (total activity) did not vary greatly, mean values of 0.51 +/- 0.04 mM. Values derived for the inactive enzyme were almost identical. In contrast, Km values for active glycogen synthase in foetal livers during gestation were significantly higher than those for adult liver. Highest values were seen at day 19 of gestation (1.84 +/- 0.08 mM) followed by a steady fall to 0.55 +/- 0.05 mM in the newborn compared with a mean value of 0.48 +/- 0.04 mM for adult liver. Existence of a reduced affinity of active glycogen synthase for UDPglucose must be recognized when assaying the enzyme in foetal liver, particularly when extrapolating values to rates of glycogen synthesis in vivo. Data were obtained only after removal of an amylase-like contaminant from foetal liver samples which invalidated the radioassay of glycogen synthase. This work illustrates the care needed in the analysis of foetal tissue and the interpretation of resulting data when utilizing methods developed for adult tissue. 相似文献
15.
E Gregoraszczuk 《Endocrinologia japonica》1991,38(3):229-237
The first objective of this research was to define the capacity of corpora lutea of pig to secrete estradiol in the presence of an androgen substrate which was testosterone. The second objective was to define the synergism between gonadotropic hormones such as LH, FSH, and PRL and testosterone as measured by estradiol and progesterone secretion by two types of porcine luteal cells. Luteal cells were collected from newly forming corpora lutea (0-3 days after ovulation) and from mature corpora lutea (8-10 days after ovulation). After dispersion, luteal cells were suspended in medium M199 supplemented with 10% of calf serum and grown as monolayers at 37 degrees C. Control cultures were grown in medium alone while other cultures were supplemented with either testosterone alone at a concentration of 1 x 10(-7) M or with 10, 100, 500 ng LH plus testosterone, 10, 100, 500 ng FSH plus testosterone or 10, 100, 500 ng PRL plus testosterone. After 2 days of cultivation all cultures were terminated and media were frozen at 20 degrees C for further steroid analysis. Testosterone added to the culture medium in the absence of gonadotropins was without effect on estradiol and progesterone secretion by luteal cells collected in the corpora lutea of the early luteal phase. On the other hand testosterone added to the medium significantly increased progesterone and estradiol secretion by cultured luteal cells collected in the midluteal phase of the cycle. No additive stimulatory action of gonadotropins and testosterone on progesterone secretion was observed in cultures of luteal cells from the early luteal phase but this was not the case in cultures of luteal cells from the midluteal phase.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
16.
Hepatocyte cultures derived from 15-day foetal rats produce the liver form of pyruvate kinase (EC 2.7.1.40) only after 3 days of culture. The appearance of the liver form of the enzyme can be blocked by the addition of 5-bromodeoxyuridine on day 2 of culture, but not by addition on day 3 of culture. The reversibility of the action of 5-bromodeoxyuridine was shown when the inhibitor was added on day 2 and removed on day 4. By day 6 of culture the liver form of pyruvate kinase was detectable. The specificity of the action of 5-bromodeoxyuridine was monitored by following changes in the closely related embryonic form of the enzyme as a control. This was unaltered by the inhibitor. 相似文献
17.
The development of phenylalanine hydroxylase in rat liver; in vivo, and in vitro studies utilizing fetal hepatocyte cultures 总被引:2,自引:0,他引:2
George C.T. Yeoh Edward Edkins Kirsten Mackenzie Stephanie Fuller Julian F.B. Mercer Hans-Henrick M. Dahl 《Differentiation; research in biological diversity》1988,38(1):42-48
Phenylalanine hydroxylase (PAH) is first detected in the liver of 21-day-gestation rats. Activity increases after birth, and in 10-day-postnatal rats it is about equal to that observed in the adult. The developmental pattern for the enzyme is reflected in the level of its mRNA determined by hybridization to 32P-cDNA, which is specific for PAH. Studies with cultured adult hepatocytes reveal that the addition of dexamethasone and dibutyryl cAMP to the medium maximizes the yield of enzyme. Hepatocytes derived from 21-day-gestation rats will produce enzyme in cultures maintained in medium supplemented with dexamethasone and dibutyryl cAMP. However, less mature cells, taken from 19-day-gestation rats do not produce measurable levels of enzyme activity. The relative amounts of PAH mRNA in the respective cultures reflect the level of PAH activity. Interestingly, after 3 days of culture, 19-day-gestation hepatocytes can be shown to express PAH mRNA. Therefore, with respect to the expression of PAH, we conclude that 19-day-gestation liver cells will differentiate during culture. 相似文献
18.
The influence of the medium collected from cultured rat Sertoli cells on the spontaneous and LHRH-stimulated release of gonadotropins by incubated rat pituitary halves was examined. The homogeneity of the cultured population of Sertoli cells taken from 20-day-old rats ranged up to 98%. The cells in culture responded to FSH stimulation with characteristic morphological changes and with increased secretion of estradiol-17 beta. The hemi-pituitaries obtained from sexually mature male rats were incubated for 5 hours in the presence of Sertoli cell culture medium (SCCM) or its fractions obtained by use of ultrafiltration. The SCCM fraction deprived of MW less than 10 kD compounds exhibited a typical inhibin-like activity, whereas crude SCCM as well as its low-molecular-weight fraction stimulated the basal FSH release to about 150% and 175% of the control values, respectively. These fractions exerted an inhibitory effect on the LHRH-stimulated secretion of both LH and FSH. It is concluded that Sertoli cells cultured in chemically defined medium release, apart from inhibin, a non-steroidal, heat-labile substance of MW less than 10 kD which stimulates the basal secretion of FSH and LH and inhibits the LHRH-stimulated secretion of both gonadotropins from incubated rat hemi-pituitaries. 相似文献
19.
Activation of bovine and baboon primordial follicles in vitro 总被引:5,自引:0,他引:5
Mammalian ovaries contain a large pool of non-growing, primordial follicles. The ability to initiate growth of this pool of resting follicles in vitro and to maintain follicular growth to a stage when the oocyte could be matured and fertilized would increase the reproductive potential of valuable domestic animals, endangered species and infertile women. This paper summarizes our progress to date in activating primordial follicles of cattle and baboons. Pieces of ovarian cortex, rich in primordial follicles, were obtained from fetal bovine and baboon ovaries during late gestation. Pieces were maintained in organ culture in serum-free medium containing ITS+ (insulin-transferrin-selenium-linoleic acid-BSA) for up to 20 days and at various times during culture some pieces were fixed for histological morphometry. As early as 2 days of culture, the number of primordial follicles had decreased by 88% or 55%, whereas the number of primary follicles had increased 2.5- or 5-fold, compared to tissue freshly isolated from bovine or baboon ovaries, respectively (P < 0.01). In baboon cortical pieces a small number of secondary follicles developed during a 20-day culture period. The development of primary and secondary follicles was accompanied by an increase in diameter of both the granulosa cell layer and the oocyte. The addition of FSH (1, 10, or 100 ng/ml) had no effect on the development of follicles in bovine cortical pieces after 7 or 14 days of culture, relative to control cultures without FSH. These results show that a high percentage of primordial follicles from cattle and baboons can be activated to grow in serum-free medium in the absence of gonadotropins. Conditions that will support further growth in vitro of follicles from these species remain to be elucidated. The culture system we have developed could be used to develop such conditions and to explore factors that regulate the movement of primordial follicles into the pool of growing follicles. 相似文献
20.
The luteotropic activity of ovine placental lactogen (oPL) on different days of gestation in ewes was assessed using in vitro methods. Corpora lutea (CL) harvested on Days 45, 70, 95, 120 and 135 of gestation and during parturition were enzymatically dispersed and plated on multiwell plates. After 48 h of incubation, all cultures were terminated and media were frozen for further steroid analysis. Cells were cultured in control medium, with addition of oPL alone, or in combination with PGE2 or PGF2alpha. Supplementation of culture media with oPL increased basal progesterone secretion by cells isolated on Days 45 and 70 of gestation. There was no effect on progesterone secretion by cells isolated on other days of gestation; PGE2 added to the culture media increased progesterone production only by cells isolated on Day 70 of pregnancy. Simultaneous oPL treatment with PGE2 had a statistically significant and stimulatory effect on progesterone production by luteal cells collected on Days 70 and 95 of pregnancy. In contrast, PGF2alpha alone in culture media decreased progesterone secretion by cells isolated on Days 45, 70 and 95 of gestation, while oPL plus PGF2alpha on Days 70 and 95 of gestation protected against luteolytic action of PGF2alpha. The results showed 1) a direct effect of the oPL on luteal cells isolated on Days 45 and 70 of gestation; 2) synergism between PL and PGE2 in progesterone production; by cells isolated on Day 70; 3) and a luteoprotective effect of oPL against the luteolytic action of prostaglandin F (PGF2alpha) observed on Days 70 and 95 of gestation. 相似文献