Erythroid precursors cultured from adult mice are sensitive to H2O2 toxicity

Summary The growth of late erythroid precursors (CFU-Es) from adult bone marrow is inhibited when Iscove's modified Dulbecco's medium supplied in liquid form is used. Catalase and other H₂O₂ destroying compounds restore the capacity of culture medium to support colony development. However early precursors from adult bone marrow and fetal liver CFU-Es were resistant to H₂O₂. This work was supported by grants from the Centre National de la Recherche Scientifique, the Institut National de la Santé et de la Recherche Médicale and the Fédération Nationale des Centres de Lutte contre le Cancer.     

Organ culture of suckling rat intestine: Comparative study of various hormones on brush border enzymes

Summary Jejunal mucosa of 6 d-old rats were cultured for 24 and 48 h in the presence of thyroxine, insulin, pentagastrin, glucagon, epidermal growth factor (EGF) or dibutyryl-A-3:5-MP cyclic with or without dexamethasone (DX). The enzymes were assayed on the purified brush borders. The various agents added alone to the basic culture medium had no effect with the exception of DX on the levels of enzyme activities. Dexamethasone alone induced sucrase, stimulated maltase, and protected other brush border enzyme activities (aminopeptidase, lactase, and alkaline phosphatase). When added to DX-supplemented medium, only the following factors modified the levels of enzymatic activities observed with DX alone. Insulin (10⁻⁶ M) increased maltase, alkaline phosphatase, and lactase activity to a greater extent than DX at 24 h culture, the effect being maintained at 48 h on alkaline phosphatase only. At 48 h culture, both EGF (10⁻⁸ M) and dbcAMP (10⁻³ M) decreased DX-induced sucrase activity. The latter agent also depressed DX-stimulated aminopeptidase activity. This work was supported by the Institut National de la Santé et de la Recherche Médicale, Centre National de la Recherche Scientifique, and a grant 79.7.1243 from the Délégation Générale a la Recherche Scientifique et Technique. P. M. S. is a recipient of a grant from Fondation de la Recherche Médicale (France).     

Growth of preadipocyte cell lines and cell strains from rodents in serum-free hormone-supplemented medium

Summary Ob17 is a clonal cell line isolated from the epididymal fat pad of C57 BL/6J ob/ob mouse that differentiates into adiposelike cells in serum-supplemented medium. In serum-free medium, this cell line shows increased growth under the addition of insulin, transferrin, fibroblast growth factor (FGF), and a factor present in extract of rat submaxillary gland (SMGE). This medium is referred to as 4F. Epidermal growth factor or nerve growth factor cannot replace SMGE, whereas partially purified platelet extract can substitute for FGF but only partially for SMGE. 4F Medium is able to support the proliferation of cells from other established preadipocyte clonal lines, HGFu and 3T3-F442A, and also of preadipocyte cells isolated from the stromal-vascular fraction of rat and mouse adipose tissues. In each case 4F medium is insufficient to support the differentiation of these cells into adipocytes. Ob17 cells grown and maintained in serum-free hormone-supplemented medium retain the ability to convert to adiposelike cells after serum addition. This serum requirement for differentiation cannot be substituted by the addition of growth hormone or of other putative adipogenic factors, or both. The results are discussed with respect to the requirements for growth and differentiation of the 3T3-L1 and 1246 preadipocyte cell lines previously described. This work was supported by the “Centre National de la Recherche Scientifique” (Grant 1208-Biochimie du Développement and Grant 4162-Endocrinologie), by the “Ministère de la Recherce et de la Technologie” (Grant 81-L-1322), by the “Fondation pour la Recherche Médicale,” by NATO (Grant 1704), and by the “Institut National de la Santé et de la Recherche Médicale” (Grant 827006).     

Effect of sodium butyrate on the hepatoma cell cycle: possible use for cell synchronization

Exposure of HTC cells to sodium butyrate caused inhibition of growth. The site of growth inhibition was studied by time-lapse cinematography and [3H]thymidine incorporation studies. Evidence is presented that sodium butyrate affected the cell cycle at a specific point immediately after mitosis. Inasmuch as it does not modify the interphase duration after its removal, butyrate may be used for HTC synchronization.     

Comparative use of fructose and glucose in human liver and fibroblastic cell cultures

Summary The effect of fructose as a substitute for glucose in cell culture media was investigated in human skin fibroblast and liver cell cultures. Cells were grown for between 2 and 10 days in identical flasks in four different media, containing 5.5, mmol·1⁻¹ and 27.5 mmol·I⁻¹ glucose and fructose, respectively. In the presence of fructose, cell growth was stimulated, but less in liver cells than fibroblasts. At Day 6, increases were observed in [³H]thymidine incorporation, protein levels, and amino acid consumption, and a reduction was noted in ATP levels. In media containing 5.5, mmol·1⁻¹ glucose or fructose, consumption of fructose was four times lower than that of glucose at Day 3 and did not rise until Day 6. In fructose media, the lactate production was very low (four to five times less than that of glucose) and the pH values were always higher. Some findings were different for the fibroblasts and liver cells, owing to the specific characteristics of these two cell types in culture; this applied especially to the effects of glucose and fructose concentrations of 27.5 mmol·1⁻¹. Several possible explanation for the stimulation of cell growth in fructose medium were discussed. This work was supported by grants for the Institut National de la Santé et de la Recherche Médicale (ATP 82-79-114) and the Unité d'Enseignement et de Recherche, Le Kremlin-Bicêtre, Université Paris-Sud (C. R. 848).     

Changes in proteoglycans of cultured pig aortic smooth muscle cells during subculture

Summary Smooth muscle cells were cultured from pig aorta. Changes in both the growth and the properties of sulfated proteoglycans were observed during passage. The population doubling time during log phase growth was 34 h from Passages 3 to 7–8 but 20 h at the Passage 11, and the cell density at the stationary phase, was 86 000 and 136 000 cells/cm² at Passages 3 and 11, respectively. Structural characteristics of sulfated proteoglycans secreted into the medium were investigated after metabolic labeling with [³⁵S]-sulfate. Significant differences were observed with age in vitro: a) [³⁵S]proteoglycan complexes were in a greater amount at Passage 10 than at Passage 3; b) the hydrodynamic size of at least 45% of subunits and about 90% of monomers decreased with in vitro aging; c) this decrease in the size of proteoglycans was partly due to a decrease in the size of their glycanic chains; d) an increase of 15% in the proportion of dermatan sulfate was observed when cells were subjected to 10 passages. This work was supported by grants from the Institut National de la Santé et de la Recherche Médicale (INSERM, U. 181) and the Fondation pour la Recherche Médicale.     

Regulation of vascular development by fibroblast growth factors

Fibroblast growth factors (FGFs) are potent stimulators of angiogenesis in vitro and in vivo. However, the precise role of FGFs and vascular development in normal and pathological tissue has long remained ill defined. Recently, substantial progress has been made toward a better understanding of their role. Genetic studies in mice or in culture systems indicate a role for FGFs in vessel assembly and sprouting. FGFs also stimulate blood vessel branching and lymphangiogenesis. The molecular mechanisms by which FGFs mediate angiogenesis are also better understood. Finally, the FGF/FGF-receptor system has become a focus for the development of novel therapeutic strategies for the treatment of angiogenesis-related diseases such as tissue ischemia.Work described herein from our laboratory was supported by grants from the Ligue Nationale contre le Cancer, the Association de la Recherche sur le Cancer, Rétina France, the Institut National de la Santé et de la Recherche Médicale (INSERM), and the Ministère de la Recherche     

Growth in serum-free medium of human colonic adenocarcinoma cell lines on microcarriers: A two-step method allowing optimal cell spreading and growth

Summary Human colonic adenocarcinoma cells have been successfully grown on polystyrene microcarriers by modifying the culture conditions used in monolayer culture. The method can be divided into two culture phases: a) a phase of spreading, wherein cells were seeded in presence of serum-supplemented medium; b) a phase of active growth wherein spread cells on the beads were allowed to grow in a serum-free medium. Under these conditions, optimal spreading and growth of HT 29 and HRT 18 cells on the microcarriers were obtained. A differential propagation was observed between HT 29-D4 and HT 29-D9 cells (both clonal populations derived from HT 29 cells) on the microcarriers that is tentatively related to the discrepancy observed in the spreading efficiency of these clonal cells on serum-coated culture flasks. An index of spreading efficiency (IS index) has been defined to quantify the efficiency of spreading of each cell line on microcarriers. These data gave the opportunity to develop serum-free, scale-up methods to culture cells like HT 29 which release potentially useful products. This work was supported by CNRS (U.A. 202 and U.A. 1186), Fédération Nationale des Centres de Lutte Contre le Cancer (FNCLCC), INSERM (CRE, n^o 847006), CNAMTS-INSERM (8386), MRT (GBM 85M0564) and l'Association pour la Recherche sur le Cancer (ARC 86-234).     

Laminin provides a better substrate than fibronectin for attachment,growth, and differentiation of 1003 embryonal carcinoma cells

Summary Culture of cells in hormonally defined media has allowed (a) the demonstration of physiological responses from cells usually unable to express them in vitro and (b) the study of the effects on growth and differentiation of diffusible factors and attachment factors. The embryonal carcinoma line 1003 forms multidifferentiated tumors in vivo but is unable to differentiate in vitro when grown in serum-containing medium. In a defined medium containing insulin, transferrin, selenium, and fibronectin as attachment factors, 1003 cells grow for several generations and differentiate into neurons and embryonic mesenchyme (Darmon et al., 1981, Dev. Biol. 85: 463–473). In the present work the effects of fibronectin and laminin were compared. In the presence of laminin the cells attached and spread better, grew faster, and could be plated at lower densities. Neurite extension was also better under these conditions and most importantly, it was found that laminin induced an important formation of muscular tissue when the cells had been seeded at low densities. Multinucleated myotubes could be stained with antibodies directed against embryonic muscular myosin. Coating the dishes with polylysine or adding FGF or serum-spreading factor to the medium allowed growth of low-density cultures with fibronectin instead of laminin but muscular differentiation was not detected under these conditions. Addition of fibronectin to laminin-containing medium did not inhibit muscular differentiation. Presented in the symposium on Plant and Animal Physiology in Vitro at the 33rd Annual Meeting of the Tissue Culture Association, San Diego, California, June 6–10, 1982. This research was supported in part by grants from the “Centre National de la Recherche Scientifique” (LA 269), the “Délégation Générale à la Recherche Scientifique et Technique,” the Fondation pour la Recherche Médicale Fran?aise,” the “Institut National de la Santé et de la Recherche Medicale,” the “Ligue Nationale Fran?aise centre le Cancer,” and the “Fondation André Meyer.” This symposium was supported in part by the following organizations: Bellco Glass, Inc., California Branch of the Tissue Culture Association, Collaborative Research, Hana Media, Hybridtech, K C Biological, Inc., and Millipore Corporation.     

Recherches sur la cuticule

Sans résumé Avec 11 figures dans le texte Une partie de ces recherches a été effectuée à l'aide d'instruments prêtés par le “Fonds National Belge de la Recherche Scientifique”. Cette liste ne comporte que les mémoirescités dans le texte. Quelques-uns ne me sont connus que de seconde main; leur indication est prédédée d'un^*.     

Isozyme differentiation of aldolase and pyruvate kinase in fetal,regenerating, preneoplastic,and malignant rat hepatocytes during culture

Summary Aldolase and pyruvate kinase isozymes were investigated in cultured hepatocytes from fetal, regenerating, and 2-acetyl-aminofluorene-fed rat liver as well as in some epithelial liver cell lines. Our results show that: (a) cell proliferation and prolonged expression of specific isozymes were found only in cultured hepatocytes from 17-day old fetuses; (b) the fetal type of pyruvate kinase expressed in regenerating and carcinogen-treated liver was temporarily lost only in cultured hepatocytes from regenerating liver; (c) the adult type of aldolase and pyruvate kinase was absent in one epithelial cell line derived from a carcinogen-treated liver and in the hepatoma tissue cell (HTC) line but was found in the Faza clone of the Reuber H₃₅ cell line during the 50 first passages in vitro; and (d) the isozyme pattern of pyruvate kinase was always more strongly shifted than that of aldolase. The observations suggest that: (a) hepatocytes from carcinogen-treated liver exhibit the same lack of ability to proliferate in primary culture as normal adult hepatocytes; (b) adult hepatocytes can produce fetal isozymes without prior cell division; (c) pyruvate kinase is a stronger marker of dedifferentiation (retrodifferentiation) than aldolase; and (d) regulatory processes of isozyme expression are different during ontogenesis, regeneration, and hepatocarcinogenesis. This work was supported by the “Institut National de la Santé et de la Recherche Médicale” and the “Fondation pour la Recherche Medicale Fran?aise”     

Ability of mammalian fibroblasts to grow in synthetic medium containing neither serum nor exogenously added macromolecules

Summary Rous sarcoma virus transformed Chinese hamster fibroblasts, clone CHR1-3, were established at high temperature, then subcloned. Six subclones with round and flat morphology harboring undeleted and partially deleted RSV proviruses, respectively, were seeded into serum-free synthetic medium with no macromolecular additives, and maintained for 2 mo. One flat subclone no. 14, fully designatedsfCHR1-3.14 for itsserum-free phenotype, was further propagated in the same medium. The cells grew exponentially in loosely attached monolayers and dould be serially passaged on bare polystyrene with an average population doubling time of 46 h. Cell attachment could be improved by using collagen-coated polystyrene or by adding a methionine supplement to the culture medium. Furthermore, thesfCHR1-3.14 cells could be subcloned and further grown in nonselective medium. The reversion rate of thesf phenotype was estimated to be 1 to 2%/cell generation. Evidence for an autocrinal stimulation was obtained by cloning efficiency assays showing a requirement for a threshold cell density. Slight growth stimulation could also be detected in assays using conditioned medium fromsfCHR1-3.14 cells and serum-restrictedwild-type (wt)NIH3T3, but notwtCHR1-3.14, cells as indicator cells. Finally,wtNIH3T3 cells used in these assays were assayed for serum-free growth and found to be able to develop their ownsf phenotype; in this respect they resemble the previously establishedsfCHR1-3.14 cells. Supported by grants from CNRS, INSERM, contract 852012, Association pour la Recherche sur le Cancer, and Fondation pour la Recherche Médicale. V. K. was supported by the Association pour la Recherche sur le Cancer and by the Fédération Nationale des Centres de Lutte contre le Cancer.     

Characterization of a new cell line,XL2, obtained fromXenopus laevis and determination of optimal culture conditions

Summary A new amphibian permanent cell line is described. It is called XL2 and was initiated from Stage 35 tadpoles ofXenopus laevis. The cell line has an epithelioid morphology and most cells can be classified into two populations with respective chromosome modal numbers 36 and 74. Contact inhibition is low. Its growth is vigorous in L15 or MEM medium supplemented with fetal bovine serum. The mean doubling time is 39 hr and the saturation density is 700,000 cells/cm² at 25°C. The absolute plating efficiency is about 70%. Cell line XL2 is unable to grow in L15 medium containing a macromolecular fraction of fetal bovien serum. Growth is restored if the latter medium is supplemented with 10 μg/ml of hypoxanthine. Optimal conditions for the dye exclusion test, for harvesting the cells, and for cloning in petri dishes are described. This work was supported by Grant 3,4514,79 of the Fonds de la Recherche Scientifique Médicale (Belgium) and by a grant of the Fonds de Développment Scientifique of the Catholic University of Louvain. The authors are indebted to Mrs. M. S. Denis, Mr. F. Desneux, and Ms. J. Janssens for competent technical assistance. Smith Kline-RIT, Genval (Belgium), is acknowledged for the generous gift of antibiotics and for performing the cultures for mycoplasma detection.     

Established cell lines of mouse marrow adherent cells producing differentiation factor(s) for the granulocyte-macrophage lineage

Summary Two continuous cell lines derived from long-term cultures of AKR mouse bone marrow adherent cells were isolated. These cell lines release colony stimulating activity (CSA), a factor that induces in vitro differentiation of granulocyte-macrophage progenitor cells. The colony forming cells and cluster forming cells in mouse marrow responsive to CSA from cell line conditioned medium were compared with those responsive to CSA from mouse lung conditioned medium (MLCM). Colony forming cells were characterized by analysis of their density distribution after equilibrium centrifugation in density gradient. Cluster forming cells were characterized by analyzing the progeny of individual clusters after transfer to fresh semisolid culture medium containing MLCM. The results obtained indicate that the CSA from cell line conditioned medium closely compares with the CSA from MLCM in terms of the populations of colony and cluster forming cells stimulated. This work was supported by a research grant from the Institut National de la Santé et de la Recherche Médicale (CRL 802620), Paris, France.     

Déclenchement en serre d'une épizootie aEntomophthora fresenii surAphis fabae par introduction d'inoculum et régulation de l'humidité relative

Résumé Deux expérimentations ont été faites en serre, dans le but de déclencher des épizooties àEntomophthora surAphis fabae Scop. par dissémination de pucerons vivants, infectés parE. fresenii Nowal. dans des colonies saines et par brumisation. Un relèvement moyen de l'humidité relative ambiante, obtenu par brumisation, même assez faible (10%), est suffisant pour avancer la saturation nocturne et retarder la baisse matinale de l'humidité relative. Ceci, ainsi que le maintien d'eau liquide sur les plantes pendant une grande partie de la journée, a permis le déclenchement des processus épizootiques. Durant la 1^re expérimentation, la croissance des colonies est arrêtée 13 j après la distribution de l'inoculum et les populations sont détruites à 80% 27 j après cette distribution. Durant la 2^e expérimentation la croissance des colonies est arrêtée 6 j après la distribution de l'inoculum et les populations sont détruites à 90% 24 j après cette distribution. Les meilleurs résultats (2^e expérimentation) ont été obtenus avec une serre complètement fermée et un inoculum distribué très peu de temps après l'installation des pucerons. Les r?les respectifs de l'eau libre et de l'humidité relative ambiante sur l'épizootiologie du champignon sont discutés, ainsi que les problèmes posés par l'influence de la brumisation sur les champignons phytopathogènes, par l'action des fongicides sur lesEntomophthora, et par la compatibilité de telles méthodes de lutte avec les traitements insecticides.

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Summary Two experiments were carried out in a glasshouse, in order to control the populations ofA. fabae on broad beans by dissemination of living infected aphids (E. fresenii) in the healthy colonies, and enhancement of the relative humidity by means of brumization. Sprinkling was released by an electronic detector of moisture. During the 1st experiment the detector ran permanently, in the 2nd experiment from 4h p.m. to 10h a.m. only. The results showed that the induced increase of the relative humidity through sprinkling was weak (10%) but enough to carry the nocturnal saturation forward and to delay it during the morning. During the 1st experiment (16th May–12th June 1978) the colony growth was arrested 13 days after the spread of the infected aphids had occurred and the proportion of dead aphids reached a high of 80%, 27 days after the spreading of the infected ones. During the 2nd experiment (18th October–13th November) the growth of the colonies was arrested in the treated plot from the 6th day after the spreading of the infected aphids and the proportion of dead aphids exceeded 90% 24 days after the spreading of the infected ones. In the untreated plot the highest level of population was three fold the one in the treated plot. The best results (2nd experimentation) were obtained when the glasshouse was completely closed and when the inoculum (infected aphids) was spread a very short time (2 days) after the infestation of the beans by the healthyA. fabae. Many questions are stated, especially concerning the role of water covering the beans and aphids on the epizootiology, the influence of sprinkling on the plant fungal diseases, the influence of fungicides on theEntomophthora or the best way to spread the inoculum. The solution may be to consider theEntomophthora as a component of integrated control in glasshouses.

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Avec la collaboration technique d'Augustine Gellé.     

Étude chromatographique des acides phénoliques et des phénols obtenus par la fusion alcaline des acides humiques

Resumé Les résultats de cette étude portent principalement sur l'analyse chromatographique des acides aromatiques et phénols, obtenus après la fusion alcaline, des acides humiques, de la lignine, de mycélium de microchampignons, de la ligno-protéine, du DNA et du RNA. Par chromatographie sur papier 70 substances sont mises en évidence dont une vingtaine sont identifiées. Les acides nucleiques mis à part, l'analyse chromatographique donne des résultats pratiquement identiques pour tous les complexes éxaminés. On démontre également que l'acide 3.5-OH-Benzoique n'est pas uniquement d'origine microbienne. Par chromatographie en phase gazeuse, le phénol, o-crésol et p-crésol sont détectés dans les acides humiques, tandis que dans la lignine le m-crésol est mis en évidence en plus de trois substances citées. Recherches subsidiées par l'Institut pour l'Encouragement de la Recherche Scientifique dans l'Industrie et l'Agriculture (IRSIA)     

Variabilité de la sensibilité despodoptera littoralis [Lep.: Noctuidae] a l'hyphomycete entomopathogèneNomuraea rileyi

Résumé La variabilité de la sensibilité des larves de la noctuelle égyptienne du coton,Spodoptera littoralis Boisduval à l'hyphomycète entomopathogèneNomuraea rileyi (Farlow) Samson a été abordée en étudiant la variabilité interclonale du champignon et la variabilité de l'insecte-h?te à travers des populations d'origines géographiques différentes. En raison de la polyphagie de l'espèce-cible, l'influence de la plante-h?te sur la sensibilité des larves phytophages a aussi été prise en compte. Les traitements ont consisté à exposer les larves à un inoculum sporal déposé par pulvérisation sur des rondelles de feuille calibrées pendant 24 h. L'activité pathogène, à l'égard des larves du 1^er stade, des 12 clones monosporaux de la soucheN. rileyi 5 varie significativement (TL50 compris entre 4,7 et 6,2 j à la dose de 3.10⁴ spores/cm²). Cette variabilité intraspécifique observée à un moment donné est néanmoins du même ordre que la variabilité de l'activité de la souche-mère polyclonale, relevée sur une période de 4 ans (TL50 compris entre 4,5 et 6,6 j). Les 3 populations originaires d'Isra?l, d'Egypte et du Maroc ont présenté une sensibilité comparable àN. rileyi 5 après contamination des larves nouveau-nées avec des doses croissantes d'inoculum (DL50 comprises entre 0,7.10⁴ et 1,1.10⁴ spores/cm² et TL50 compris entre 5,1 et 5,5 j à la dose de 3.10⁴ spores/cm²). L'alimentation des larves, depuis l'éclosion, sur 4 espèces végétales: chou, coton, féverole ou aubergine, n'a pas modifié significativement la sensibilité des larves du 3^e stade (DL50 comprises entre 0,3.10³ et 0,7.10³ spores/cm² et TL50 compris entre 4,0 et 4,3 j à la dose de 3.10⁴ spores/cm²). La souche polyclonaleN. rileyi 5 para?t bien adaptée à la noctuelle égyptienne du coton en raison de sa stabilité, de son agressivité à l'égard des populations d'origines géographiques différentes et de l'absence d'effet de l'alimentation des larves sur l'infection.      

A brief history of genome research and bioinformatics in France

The development of in silico genomics has progressed slowly in France for a number of political reasons. Two administrative organizations, the Groupement de Recherche sur les Génomes (GREG) and the Groupement de Recherche 1029 (GDR 1029) of the Centre National de la Recherche Scientifique (CNRS) have been established. These organizations have created the dynamics that hopefully will place France (which coordinated consortia that completed several of the first large microbial genomes) among the developed nations that support Large-Scale Biology.     

Zytophotometrische Bestimmung des Histon- und Gesamtproteingehalts von Zellen des lymphatischen Gewebes

Résumé L'étude au microscope électronique a révélé la présence de particules B dans le thymus de la souris NZB, adulte et saine. Les particules se forment dans un type de cellule thymique bien défini, la cellule réticulo-épithéliale à vacuoles et naissent par bourgeonnement des membranes plasmatiques limitant les vacuoles. L'élaboration de particules B dans lethymus d'animaux apparemment sains est un fait nouveau dont la signification n'est pas encore connue.

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Summary An electron microscopical study revealed the presence of virus-like particles in the thymus of adult healthy NZB-mice. The particles are found in the vacuoles of reticuloepithelial cells and are elaborated by a budding process from the membranes surrounding the vacuoles. The formation of B-particles in the thymus of apparently healthy animals is a so far unknown fact the significance of which remains to be determined.

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Avec l'aide du Centre National de la Recherche Scientifique et de l'Institut National de la Santé et de la Recherche Médicale, et la collaboration technique dévouée de M^lle J. Patry et de Mr. B. Fontaine.     

A method for separating cultured preadipocytes according to their density: Application to stromal cells from overfed suckling rats

Summary The stroma vascular fraction of adipose tissue consists of a heterogeneous cell population; not all the cells in this compartment undergo adipose conversion in primary culture. A density gradient centrifugation procedure was used to separate cultured cells on the basis of their triglyceride content. This method was applied to both stroma vascular cells from rat adipose tissue and to a 3T3 F442A preadipose cell line as a reference. Comparison of the results obtained from these two cell types suggests that this separation procedure can lead to a quantification of adipose differentiation in the heterogeneous stroma cell population. Separation procedures were applied to cultured stromal cells derived from young rats during the onset of nutritional obesity induced by overfeeding in early life. Results show that early overfeeding induced an increase in the stromal cell differentiation capacity which is expressed in vitro. This work was supported in part by Institut National de la Santé et de la Recherche Médicale (CRL n^o 82-70-22).