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Differences in insulin-like growth factor binding proteins (IGFBPs) expressed within estrogen receptor positive (ER+, MCF-7/6) and negative (ER-, MDA-MB-231) human breast cancer cells cultured in chemically defined medium were observed. In the absence of insulin, 17 beta-estradiol affects this expression in ER+ cells by significantly reducing 34 and 28 kDa species. In ER+ cells, insulin appears to minimize the estrogen induced reduction of these 34 and 28 kDa IGFBPs and stimulates a 24 kDa type. We suggest that through its association with a given IGFBP, insulin-like growth factor-II (IGF-II) directs lysosomal enzymes to secretion by its binding to the mannose-6-phosphate/IGF-II receptors present in the Golgi apparatus. Alternatively, the association of IGF-II with another IGFBP would inhibit this binding and lead to its autocrine or intracrine mitogenic action via the IGF-I receptor.

Cytokines like tumor necrosis factor (TNF), interferon-gamma (IFN-gamma), and interleukin-1 (IL-1) are known to interfere with the differentiation of cultured cell lines of adipocyte precursors. In the present study, the effect of mouse and rat IFN-gamma, as well as human IL-1 beta, was investigated on rodent preadipocytes in primary cultures, either in the presence of fetal bovine serum (FBS, 10%) or in serum-free defined medium. IFN-gamma exerted an antiproliferative action that was more pronounced when cells reached confluency than during the growth phase of the culture. Morphological observation and quantifications of undifferentiated and differentiating cells revealed that IFN-gamma caused a decrease in the proportion of cells devoid of lipid droplets which would correspond to fibroblast-like cells, whereas preadipocytes remained unaffected. IFN-gamma induced a marked retardation of adipoconversion, resulting in a partial inhibition of lipoprotein lipase (LPL) activity and a severe decrease in glycerol-3-phosphate dehydrogenase (GPDH) activity. The antiproliferative and anti-LPL effects of IFN-gamma were neutralized by adding anti-IFN-gamma antibodies, while these antibodies prevented only partially the depressing effect of IFN-gamma on GPDH activity. Contrary to IFN-gamma, IL-1 beta slightly enhanced the proliferation in preadipocyte cultures. IL-1 beta also depressed adipoconversion, inhibited markedly LPL activity, and partially reduced GPDH activity. These results show that the influence of cytokines on adipoconversion observed in preadipocyte cell lines can be found in normal preadipocytes in culture.

When an isocaloric low protein diet (8% versus 20%) is administrated to rats during gestation, the fetus or the neonate has a lower body weight and the structure and function of the endocrine pancreas is altered: islet cell proliferation, islet size and islet vascularisation are reduced when compared with a control group. If these fetal islets are cultured during 7 days in vitro, they secrete less insulin than normal islets in response to an AA challenge. When these newborns are fed with a low protein diet until adult age and analyzed at 70 days, the same alterations of the endocrine pancreas persist: the in vitro insulin secretion of the isolated islets in response to AA is dramatically depressed although their response to glucose is normal. However in vivo glucose and insulin levels in response to oral glucose challenge are abnormal. In addition, permanent functional alterations seem to persist when induced in utero. A normal diet (20% protein) given from birth to adulthood does not restore a normal insulin response in vivo to an oral glucose challenge.

The present study was aimed at localizing gamma-aminobutyric acid (GABA) and its enzyme of synthesis, glutamic acid decarboxylase (GAD), in the mouse pancreas by immunocytochemical methods. The influence of GABA on hormone release was also studied with normal mouse and rat islets and the isolated perfused rat pancreas. Particular attention was paid to glucagon release to test a recent hypothesis suggesting that GABA mediates the still unexplained glucose-induced inhibition of glucagon release. GABA and GAD were identified only in islet cells and never in the exocrine tissue. Exogenous GABA, baclofen (agonist of GABAB receptors), muscimol (agonist of GABAA receptors), or bicuculline (antagonist of GABAA receptors) did not affect insulin and somatostatin release by isolated mouse or rat islets. GABA was also without effect on glucose-induced electrical activity in mouse B-cells. Glucagon secretion by mouse islets was only slightly inhibited (approximately 20%) by GABA. Since muscimol had a similar effect, and baclofen was ineffective, the inhibition by GABA probably involves GABAA receptor activation. Bicuculline, however, did not antagonize the inhibitory effects of GABA and muscimol, probably because the antagonist alone also decreased glucagon secretion. In contrast to GABA, low (3 mM) and high (20 mM) concentrations of glucose strongly inhibited (approximately 50-65%) glucagon release; this inhibition was not prevented by bicuculline. Similar results were obtained with the perfused rat pancreas; muscimol slightly inhibited glucagon release under various conditions, and bicuculline did not reverse the strong inhibition produced by 16.7 mM glucose. In conclusion, GABA does not affect insulin and somatostatin secretion, but inhibits A-cells, probably by acting on GABAA receptors. It is unlikely, however, that this small inhibitory effect can account for the inhibition of glucagon release produced by glucose.

The effects of physiological glucocorticoids such as cortisol and corticosterone, as well as dexamethasone, on proliferation and differentiation of rat fat cell precursors kept in primary culture were analyzed. In serum-containing medium (10%), glucocorticoids markedly decreased cell proliferation, either on subconfluent or on confluent cultures. This effect was independent of the presence of insulin. In contrast, acute amplification of adipose conversion was observed mainly when glucocorticoids and insulin were added simultaneously. Morphological quantification of lipid-containing cells confirmed acceleration of the maturation process, and an early and specific reorganization of the cytoskeleton was detected at the ultrastructural level. In the presence of insulin, glucocorticoids also enhanced the main marker enzymes, lipoprotein lipase, and glycerol phosphate dehydrogenase. Glucocorticoid effects on precursor proliferation and differentiation were clearly dose-dependent, dexamethasone being 10 times more potent than cortisol and corticosterone. Similar results were obtained in serum-free medium, as well as in preadipocyte cultures derived from different fat deposits. This study demonstrates that in addition to an acute inhibition of precursor growth, glucocorticoids exert a clear stimulation of adipose conversion, which depends mainly on the presence of insulin and the glucocorticoid concentration.

After conjugation between Chlamydomonas gametes of opposite mating type, a transient dikaryon is formed. The two nuclei fuse within 4-6 hr after mating. The young diploid zygote differentiates into dormant zygospore competent to complete meiosis, or more rarely (2-10% of cases) it undergoes mitosis to produce a stable diploid progeny. We here bring genetical, biochemical, and cytological evidence that among the mitotic zygotes, a large proportion of them undergo cytokinesis without fusion of the nuclei-a process that has been termed "cytoduction." By using appropriate genetic markers, haploid cytoductants that possess the nuclear genotype of one parent and the chloroplast marker of the other parent can easily be isolated. Genetical analysis and hybridization experiments moreover show that many haploid cytoductants transmit the chloroplast DNA molecules of both parents and that, as in diploids, these DNA copies occasionally recombine. This process of cytoduction extends the life cycle of Chlamydomonas and provides new tools for its genetic analysis.

The aim of this study was to localize cells immunoreactive for glutamate decarboxylase (GAD), the enzyme of GABA synthesis, in pyloric and oxyntic regions of the rat stomach as well as in the rat and mouse pancreas. GAD immunocytochemistry was carried out on polyethylene glycol or cryostat sections of alkaline paraformaldehyde fixed tissue, with simultaneous immunolabelling of various gastro-pancreatic hormones for topographical comparison. In the rat stomach, nerve fibers displaying intense GAD-like immunoreactivity were seen in the myenteric plexus, the circular muscular layer, the submucosa and the lamina propria of the mucosa. But, they were absent from the submucous plexus. Colchicine treatment of the rats allowed to detect some labelled perikarya in the myenteric plexus suggesting that the GABAergic innervation is at least partly intrinsic to the stomach. In the oxyntic and pyloric mucosa, endocrine cells appeared immunostained for GAD. However, the nature of their hormones remained unknown since double immunodetections revealed that they were immunoreactive neither for gastrin nor for somatostatin. In the rat and mouse pancreas, GAD-like immunoreactivity was found in islet cells which corresponded only to insulin-secreting cells. Somatostatin-, glucagon- and pancreatic polypeptide-immunopositive cells were devoid of GAD immunolabelling. No GAD-like immunoreactivity was detected in the exocrine tissue and innervation. These results strenghten the hypothesis that GABA is not only a neurotransmitter in the stomach but that it could also be an endocrine or paracrine factor in the stomach and pancreas.

Cell suspensions prepared by collagenase digestion of pancreata obtained from 21.5-d-old rat fetuses were preincubated in RPMI medium containing 10% fetal bovine serum (FBS), to ensure cell adhesion. Twenty hours later, this medium was replaced by a chemically defined medium. Dulbecco's modified Eagle's (DME)-F12 was used alone or supplemented with various combinations of transferrin, sodium selenite, or Ultroser G. The evolution of the culture and the islet ultrastructure were similar in defined and serum-containing media. However, in the defined medium, the neoformed islets seemed less numerous, and the fibroblast layer less dense, when compared to the RPMI + 10% FBS control medium. At Day 7, in defined media, the total insulin content per dish was half that of control cultures. None of the tested additives improved the yield of the cultures. The fractional insulin release per day was elevated in defined media. In subsequent incubations, glucose and leucine stimulated insulin release in a way characteristic of these cells of fetal origin. The labeling index of islet cells cultured in DME-F12 reached 10.7%, which is not far from that observed in RPMI + 10% FBS. Such a defined medium is useful to study B cell physiology, avoiding the possible interaction of serum components with substances to be tested.

The linear mitochondrial DNAs of the two infertile algal species Chlamydomonas smithii and C. reinhardtii are co-linear with the exception of a 1 kb intron (alpha intron) located in the cytochrome b gene of C. smithii. C. smithii also possesses an additional HpaI restriction site (H marker) located in the COXI gene, about 5 kb from the intron. In reciprocal crosses, C. smithii (H+ alpha +) x C. reinhardtii (H- alpha -), the alpha intron is transmitted to all diploid progeny, whereas the H marker is frequently transmitted either biparentally or paternally depending on whether the C. smithii parent is maternal (mt+) or paternal (mt-). In diploids resulting from artificial fusion between vegetative cells, the absolute transmission of alpha is accompanied by the frequent transmission of the H+ marker, irrespective of the mating type of the parental strains. Finally, in reciprocal crosses between C. smithii (H+ alpha +) and recombinant H- alpha + clones, the transmission of the H marker is predominantly paternal or biparental. These results allow us to conclude that (1) the alpha intron behaves as a group I intron whose unidirectional conversion influences the transmission of the H marker; and (2) the mt- paternal mitochondrial genome is transmitted more often than the mt+. The mating type has no effect in diploids obtained by artificial fusion.

The administration of a low protein (LP, 8% protein/dry matter) but isocaloric diet to gestating rats did not affect their fertility, but slightly reduced the quantity of food intake as well as body weight gain. The LP diet also did not affect the placental weight, but the weight of the offspring was decreased. Accordingly the fetal endocrine pancreas was altered by the LP diet. Two different morphometric analyses showed that in the LP neonate B-cell proliferation and islet size were reduced in the head of the pancreas. In the pancreatic tail, these parameters were also decreased but to a lesser extend. Islet vascularization in the neonates was dramatically reduced in both parts of the pancreas when the mothers were fed with the LP diet.

The feasibility of maintaining long-term viability of human venous allografts by cryopreservation has been investigated. Segments of vein were obtained from 85 patients undergoing a stripping operation for varicose veins. The venous segments were immersed in a dimethylsulfoxide 15% solution, deep frozen at -196 degrees C in liquid nitrogen and preserved for a duration of 1 week to 24 months. Light microscopy (n = 126) failed to demonstrate striking differences between control veins and any of the cryopreserved veins. The types of damage observed at scanning electron microscopy included endothelial cell separation, endothelial cell loss, exposed basement membrane and exposed fibrillar collagen, which were graded on a scale. The score for short term (less than 3 weeks) stored veins was 8.1 +/- 0.9 (mean +/- SEM) and did not differ from the long-term (greater than 10 weeks) stored veins score (6.3 +/- 1.0, p NS). The tissue enzymes LDH, GOT, GPT, CPK were measured in the frozen vein groups (n = 115) after thawing to room temperature. Cryopreservation did not alter any of the tissue enzymes measured when compared to controls. Endothelial fibrinolytic activity (FA) of 58 venous segments cryopreserved for a mean duration of 20 months was 6136.4 +/- 292.1 Tissue Activator Units (TAU) and did not differ from FA of 11 controls (5989.1 +/- 696.8 TAU). Synthesis of 6-Keto-PGF1-alpha-2, a stable breakdown product of PGI2, measured in 10 venous segments cryopreserved for 10 months, was significantly higher than in 13 veins stored in saline for 12 hours at 4 degrees C (2.8 +/- 0.4 vs 0.4 +/- 0.1 PG ml-1mg-1min-1, respectively; p less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

The stroma-vascular fraction (SVF) of inguinal and epididymal fat pads of 4 week-old rats was studied by electron microscopy. Among the various cell types, endothelial cells and preadipocytes were found in both SVF, while mesothelial cells were only detected in the epididymal SVF. The resulting heterogeneity of primary culture and the adipoconversion of the fat cell precursors were studied in a serum-supplemented medium enriched with insulin (14.5 nM) and exogenous triglycerides. Despite the heterogeneity of the inoculum, the primary cultures were rather homogeneous, fat cell precursors being the main cell type. Distinctive contaminant fibroblast-like cells were observed in both cultures, whereas epithelial-like cells, which correspond most probably to mesothelial cells, were only found in epididymal cultures. Differentiation of fat cell precursors was assessed by the appearance of lipoprotein lipase (LPL) and glycerol-3-phosphate dehydrogenase (GPDH). LPL activity was found in the same level in cells of both deposits while GPDH activity was elevated in inguinal vs epididymal derived stroma-vascular cells. The different adipose conversion pattern of both cultures was confirmed by morphological quantification: the maturation of epididymal fat cell precursors was faster but less extensive. These differences could be related mainly to regional localization rather than to different maturation of the two fat deposits.

There are now increasing evidences suggesting that GABA is able of direct interaction with certain endocrine cells. In the present study, highly specific anti-GABA-glutaraldehyde antibodies and 3H-GABA uptake were used at the light and electron microscope levels to investigate the occurrence of cells containing endogenous GABA or taking up exogenous GABA in the mucosal antrum and corpus of the rat stomach. Only certain endocrine cell types of both regions were immunostained or grain-labelled. However, the morphology of their secretory granules did not allow to identify the nature of their hormone with certainty but suggested that somatostatin-like cells could interact with GABA. The combination of gastrin and somatostatin immunodetection with 3H-GABA uptake autoradiography at the light microscope level, revealed that a subpopulation of somatostatin-like cells and other still unidentified endocrine cells are able to take up GABA, while the gastrin-like cells are not. These results reinforce the hypothesis that certain endocrine cell types of the diffuse endocrine system of the digestive tract are able to directly interact with GABA.

Comparative ultrastructural stereology of 6 and 24-month-old rat B-cell cytoplasm revealed an increase with age in secondary lysosomes and a decrease in the volume density of RER and Golgi apparatus. The reduction of RER observed in freshly isolated islets could affect (pro)insulin biosynthesis in vitro: if the initial mobilization of precursor molecules for protein synthesis was the same, a delay was noted in their transit to the Golgi apparatus in B-cells of old islets. No further differences were seen in the autoradiographic distribution of radioactive amino-acids. More, the stock of insulin granules was similar in all age groups in both in vivo and in vitro conditions. Neither were any differences observed in the insulin secretion into culture media as well as during a subsequent incubation in supraphysiological glucose concentrations.

Autogenous mesothelium was used as venous substitute in ten dogs. Patches of mesothelium of three different origins were grafted into the anterior wall of the common iliac veins (CIV): peritoneum taken from and including the posterior rectus sheath (PRS), simple peritoneum (P) and mesentery (M). Animals were killed after 2, 4, 8, and 16 days and after 3 months. The segments of CIV, including the patches, were removed for study. On light microscopy, the PRS grafts showed a normal mesothelium but marked submesothelial fibrosis. The M and P grafts showed normal mesothelium and only mild fibrous thickening. On scanning electron microscopy, there was a perfect continuity of the mesothelial cells and the normal endothelium at the suture line. In the center of the graft, the cells had become elongated along the axis of blood flow. Fibrinolytic activity (FA) was measured by a standardized fibrin plate technique and quantitated in tissue activator units per gram of tissue (TAU/g). The mean FA of iliac vein specimens was 1101.7 +/- 133.3 TAU/g (mean +/- SEM). The mean FA determined before grafting for each kind of mesothelium was the following: PRS = 418.8 +/- 26.9 TAU/g; P = 873.0 +/- 107.1 TAU/g; M = 1142.3 +/- 91.4 TAU/g where only PRS showed values significantly lower than iliac vein mean FA (P less than 0.001). Postoperatively, the mesothelial FA, after an initial reduction, increased on day 4 and reached values significantly higher than the control values (1445.7 +/- 204.1 TAU/g tissue vs 853.1 +/- 62.3 TAU/g tissue; P less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

The localization of high-affinity uptake sites for 3H gamma-aminobutyric acid (3H-GABA) was investigated in the rat duodenum during ontogenesis and also at the adult stage (from 15.5 days of fetal life up to 105 days post natum) by means of low- and high-resolution autoradiography. At all stages studied, specific endocrine cell types of the epithelium were labelled and an intense uptake was detected in the nervous tissue, especially in glial cells but also in scarce neurones. When the incubation medium was supplemented with beta-alanine (1 mM), a blocker of the glial uptake for GABA, the labelling persisted only in endocrine cells and in few neurones. The intensity and the frequency of the labelling decreased at later periods compared to the earlier developmental stages. The GABA content of the duodenum as measured by a new ion-exchange column chromatography-HPLC-coupled method was higher in the early postnatal period compared to later stages. These observations suggest that GABA, in addition to being a neurotransmitter, may play an important role during development of the duodenum.

The B-cells of the endocrine pancreas constitute an adequate model for in vitro study of the aging process in highly differentiated cells. In the present study, collagenase-isolated islets of Langerhans from young and senescent rats were cultured up to 28 days. The response of the B-cells to the stimulatory conditions of the culture medium involved the nucleus, ribosomes, endoplasmic reticulum, Golgi apparatus, and secretory granules. Correlated data from light microscopy, electron microscopy, and insulin radioimmunoassay show that the differentiation and function of senescent B-cells are maintained in culture, as it has been proven for the B-cells of younger animals. On the other hand, signs of cytological deficiency not directly concerned with the specific function of B-cells were observed: abnormal mitochondria and lysosomes are more numerous in the senescent B-cells. The proliferative capacity of the B-cells of aged rats is reduced.

The occurrence of GABA-containing cells in the rat entero-pancreatic system was investigated by using anti-GABA-glutaraldehyde antibodies at the light and electron microscope level. In the pancreas, the B cells showed intense immunoreactivity, contrary to non-B and exocrine cells. Moreover, post-embedding immunogold staining was localised mostly in mitochondria, close to rough endoplasmic reticulum and in the nucleus. The insulin granules appeared nonsignificantly stained, which suggests the lack of cosecretion of GABA together with insulin. In the duodenum, GABA immunoreactivity was detected in certain endocrine cell types, suggesting a possible interaction with this amino acid. The well established GABAergic innervation in the enteric system was also confirmed by immunolabelling.

Cell suspensions prepared by collagenase digestion of pancreases obtained from rat fetuses (21.5 d old) and newborns (2.5 d old) were mixed with a collagen solution and inoculated on a collagen base layer. At the onset of the culture, most acinar cells became necrotic, whereas other epithelial cells proliferated. Most of the cell clusters arranged themselves into simple polarized structures composed of epithelial cells forming hollow spheres, and from these budded neoformed endocrine islets. Scarce fibroblasts were located close to these structures. Immunocytochemical localization of insulin and glucagon, as well as ultrastructural characteristics of the cell types revealed an intrainsular distribution similar to the in vivo localization. Tridimensional matrix of collagen offers, to perinatal pancreatic cells in culture, an environment close to the in vivo conditions: cells reorganize themselves in tissuelike structures and cell interactions concerned in the cytodifferentiation of pancreatic islets occur. This system allows for the study of undifferentiated epithelial cells--the presumed stem cells--differentiating and differentiated endocrine cells in the same preparation.

Different events such as virus infections, toxins, nutritional factors, antiinsulin antibodies may be rendered responsible for the pathological changes in the pancreatic B-cell and result in a diabetic state in the postnatal, adolescent or adult age. These different interferences which may lead to the diabetic state need full consideration and assessment if prevention and cure are to be considered.

The localization of gamma-aminobutyric acid (GABA) high-affinity binding sites was investigated in the exocrine and endocrine pancreas of neonatal rats by means of 3H-GABA autoradiography. GABA-binding was identified on Schwann cells and on the cells of the intralobular excretory ducts. In the endocrine part of the pancreas, no labelling was observed except in peripheral islet cells which, on the basis of their scarcity and distribution, could be somatostatin cells. Furthermore, peri-insular innervation showed considerable labelling.

The use of a semi-natural medium (Eagle's MEM supplemented with chicken embryo extract) appears to be satisfactory for ovarian and testicular fragment cultures of Carassius auratus. The male germ cells show a normal degeneration which might be attributed to a lack of pituitary hormones: gonadotropic and, possibly, somatotropic hormone. The ovarian fragments maintain a normal organization during 21 days of culture. Only the oocytes in late vitellogenesis massively atresiate at the beginning of culture. The addition of fetal bovine serum to this medium produces surprising effects on the testicular explants, particularly on the spermatogonial mitoses. This might be attributed to the presence of STH and androgen in the serum. The modifications of ovarian explants, on the other hand, are identical to those observed with Eagle's MEM supplemented with chicken embryo extract. M 199 has been tested as a synthetic medium; the preliminary results are encouraging. Testosterone, when added to the semi-natural medium, permits complete spermatogenesis in the testicular fragments during the 21 days of culture, but does not produce any effect on the ovarian explants.

Pancreatic islets, newly formed in vitro were incubated in the presence of streptozotocin (STZ; 0.4 mM) for up to 6 h. Ultrastructural changes first appeared between 2 and 4 h; heterochromatization, was followed by swelling of nuclear and reticular membranes, vesiculation of the Golgi apparatus, fragmentation of cell membranes and finally mitochondrial destruction. At the end of the experiment all the B cells were destroyed, whereas the other cell types remained intact. Exogenous ZnSO4 was added during preincubation periods to increase the intrainsular zinc content and to determine any protective effect against STZ-cytotoxicity. Since the addition of zinc had no obvious effect, it is suggested that STZ cytotoxicity on B cells cannot be attributed to competition for zinc between copper-zinc superoxide dismutase (Cu-Zn-SOD) and the crystallization of insulin.

Cell suspensions prepared by collagenase digestion of the pancreas of rat fetuses (21.5 days) were cultured for 7-9 days in RPMI medium containing 10 mM glucose. Exocrine cells disappeared rapidly, whereas fibroblasts and endocrine cells proliferated. These latter were first arranged in monolayers but progressively reorganized in neoformed islets essentially composed of B-cells. Total insulin content of the culture dishes increased until day 9, and fractional insulin release was about 20% per day. After 1 week, islets incubated in glucose-free medium released less than 1% of their insulin content over 2 h. Glucose (16.7 mM) caused a slower and weaker (3-fold) stimulation than 10 mM leucine or arginine (3-5-fold). The effects of the three secretagogues were potentiated by theophylline, but only those of glucose and leucine were inhibited by diazoxide. These neoformed islets thus retain a fetal character (relatively low responsiveness to glucose), but the stimulus-specificity of the inhibition by diazoxide is the same as in adult islets. This technique may be useful for studying the mechanisms which govern the organization of pancreatic endocrine cells in islets, and which underlie their functional maturation during the perinatal period.

In vitro, B-cells maintain their capacity to multiply. However, the number of mitotic figures observed in cultured rat Langerhans islets is far larger for young donnor animals than for old ones. The same observation is true for the number of nuclei incorporating tritiated thymidine in vitro. These data are in agreement with the principal observation resulting from the study of senescence on different in vitro cellular models: the decrease of the replication potential with increasing age. The stimulation of DNA synthesis by glucose, however, does not seem to be altered with aging.

Morphometrical analysis of the endocrine pancreas of senile 30-month-old rats revealed that the volume density as well as the numeric density of islets of Langerhans were much lower than in 24-month-old rats, which coincided with a much higher percentage of pycnotic nuclei in islet cells. The proportion and localization of the different categories of endocrine cells (A, B, D and PP) remained however unchanged with aging. The apparent problem of cell renewal observed in vivo in the very old age was detected earlier in vitro by tritiated thymidine incorporation. Such experiments showed that 24-month-old islet cells had a decreased labelling index when compared to 3-month-old cells. The proliferation capacity of the old cells could be partially increased by changing the serum concentration or type. Similarly as being more sensitive to serum factors, these cells underwent also more pronounced negative influence of high oxygen pressure on replication. A stereological analysis of the ultrastructure of non-degenerated B-cell nuclei revealed that with age, the relative volume of the condensed chromatin increased progressively at the expense of the dispersed form. This suggests that the still functioning senile B-cells could reduce their transcriptional activity.