共查询到20条相似文献,搜索用时 31 毫秒
1.
R Dulbecco M G Stoker 《Proceedings of the National Academy of Sciences of the United States of America》1970,66(1):204-210
Experiments were designed to discriminate between inhibition of growth due to contacts or exhaustion of serum factors. The cell layer was wounded and the migrating cells were followed by time-lapse cinematography; DNA synthesis in the same cells was recognized by means of (3)H-thymidine labeling and radioautography. In this way, the complete history of individual cells migrating to the wound could be described. The results show that topographical relationships between cells play an important role in controlling initiation of DNA synthesis. It is still unclear whether initiation is promoted by release from contacts or by the increased ability of the cells to utilize serum factors because of their changes in shapes and activities. 相似文献
2.
R. R. Bürk 《Proceedings of the National Academy of Sciences of the United States of America》1973,70(2):369-372
When a monolayer culture of normal Balb/c3T3 cells is wounded by scraping away part of the cell sheet, the cells do not migrate into the cleared area unless there is serum in the culture medium. By contrast, SV40-transformed Balb/c3T3 cells do migrate into the wound area without serum. A quantitative assay for the migration of Balb/c3T3 cells into wounds is described. This assay is used in the partial purification of a migration factor released into serum-free medium by SV28 cells. SV28 is a line of BHK21/13 hamster cells transformed by SV40 chosen for its malignancy. The most purified fractions have about 1500 times the specific activity of whole calf serum. These fractions have an activity that promotes overgrowth of Balb/c3T3 cells to high density and an activity that prolongs cell survival without serum. The SV28 migration factor is not extractable from the medium of untransformed BHK21/13 cells or from serum. This migration factor might contribute to the malignancy of SV28 cells. 相似文献
3.
R W Holley J H Baldwin J A Kiernan T O Messmer 《Proceedings of the National Academy of Sciences of the United States of America》1976,73(9):3229-3232
The growth controls observed in benzo[a]pyrene-transformed 3T3 cells (BP3T3) are compared with those of virus-transformed and normal 3T3 cells. Superficially, the chemically transformed BP3T3 cells have the same behavior as virus-transformed SV3T3 cells. Both grow to high cell density in culture medium with 10% serum, both form colonies in Methocel, and both are tumorigenic. Closer examination, however, has disclosed that BP3T3 cells exhibit "normal" growth controls at low serum concentrations. In contrast to the behavior of SV3T3 cells, the initiation of DNA synthesis in BP3T3 cells is still dependent on a serum factor. If BP3T3 cells are grown in medium with 0.2% serum, the cells become quiescent, with growth arrested in the Gu or G0 phase of the cell cycle. The addition of serum or the fibroblast growth factor (FGF) to such quiescent cells leads to the initiation of DNA synthesis and the resumption of growth. As with normal 3T3 cells, if the growth rate of BP3T3 cells is limited by a suboptimal concentration of serum, the growth rate of the cells is increased by the addition of FGF. Also, BP3T3 cells show density-dependent regulation of growth, if the medium contains a low concentration of serum. BP3T3 cells, therefore, have the behavior of "transformed" cells when cultured in medium with 10% serum, but behave as "normal" cells in medium with low serum. In comparison with normal 3T3 cells, the difference in growth behavior of BP3T3 cells appears to be due to a substantial decrease in the cells' requirement for a serum growth factor of the FGF type. Exploration of possible causes of this substantial decrease indicates that the primary cause is a lower rate of depletion of the serum growth factor from the culture medium by BP3T3 cells. The decrease in rate of depletion is sufficient to account for the uncontrolled growth of BP3T3 cells in medium with 10% serum. It is suggested that a decreased rate of depletion of a growth factor may contribute to tumorigenicity of cells in vivo. 相似文献
4.
Control of the Initiation of DNA Synthesis in 3T3 Cells: Low-Molecular-Weight Nutrients 总被引:11,自引:16,他引:11
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Robert W. Holley Josephine A. Kiernan 《Proceedings of the National Academy of Sciences of the United States of America》1974,71(8):2942-2945
Sparse 3T3 cells, in excess serum, become quiescent in the early G(1) (or G(0)) phase of the cell cycle if the cells are cultured in low concentrations of amino acids, glucose, or phosphate ion. These quiescent cells then initiate DNA synthesis if the concentration of the limiting nutrient is increased. In normal medium, DNA synthesis in the same 3T3 cell line is controlled by serum factors. The results demonstrate the complexity of the control by external agents of DNA synthesis in these "normal," density-dependent cells. 相似文献
5.
Dennis D. Cunningham Arthur B. Pardee 《Proceedings of the National Academy of Sciences of the United States of America》1969,64(3):1049-1056
Two- to fourfold increases in uridine and phosphate uptake were brought about within 10 to 15 minutes after adding fresh serum to confluent 3T3 cells. The stimulation of transport was specific, since no serum effect was observed for 3-O-methyl-D-glucose or amino acids. The early increase in RNA labeling, previously identified by (14)C-uridine incorporation, could be accounted for by this transport effect. Increased labeling with (32)P-phosphate of at least five phospholipids occurred soon after adding serum. This increase, however, could not be accounted for by increased transport. Both of these results suggest that specific early membrane changes are involved in "contact inhibition."The following results are consistent with this suggestion. Uptake of uridine and phosphate by nonconfluent 3T3 cells was higher than by confluent cells and was only slightly increased by fresh serum. In contrast, uptake of these substrates by Polyoma virus-transformed 3T3 cells, which are not subject to "contact inhibition," was not significantly decreased after the cells became confluent, and serum addition had no effect on transport by either nonconfluent or confluent Polyoma virus-transformed 3T3 cells.Fractionation of serum on Sephadex G-200 demonstrated that the factor which stimulated phosphate transport was different from the previously identified factor which stimulates DNA synthesis by confluent 3T3 cells. 相似文献
6.
Control of DNA synthesis in growing BALB/c 3T3 mouse cells by a fibroblast growth regulatory factor 总被引:8,自引:0,他引:8
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C. V. Natraj Prasanta Datta 《Proceedings of the National Academy of Sciences of the United States of America》1978,75(12):6115-6119
A cell surface component from quiescent BALB/c 3T3 mouse cells that inhibits DNA synthesis and cell division when added to a culture of growing 3T3 cells has been detected. The inhibition of DNA synthesis by this factor was dependent on concentration and time of incubation; a transient exposure of cells to the factor followed by incubation in its absence for 20 hr was sufficient to elicit its inhibitory effect. The active component appears to be protein in nature, as judged by heat inactivation and trypsin sensitivity. Extracts obtained in an identical manner from quiescent 3T3 cells that had been preincubated in situ with uridine diphosphate N-acetyl-D-glucosamine (UDP-GlcNAc) did not inhibit DNA synthesis. The effect was specific for UDP-GlcNAc: incubation with three other nucleotide sugars yielded active component. Incubation of the inactive component from UDP-GlcNAc-treated cells with purified N-acetyl-beta-D-glucosaminidase in vitro restored its inhibitory property. Extracts from growing cells failed to inhibit DNA synthesis. These results suggest that reversible glycosylation with N-acetyl-D-glucosamine residues may serve as a regulatory signal for the conversion of the active factor to its inactive form. We propose that the onset of quiescence of 3T3 cells is due to a casual relationship between depletion of growth factors in the culture medium and the presence of the active regulatory factor on the cell surface that inhibits DNA synthesis; conversion of the regulatory factor to its inactive form under favorable nutritional status may be viewed as a switch that allows DNA synthesis to resume. 相似文献
7.
Serum factor requirements of normal and simian virus 40-transformed 3T3 mouse fibroplasts 总被引:5,自引:14,他引:5
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D Paul A Lipton I Klinger 《Proceedings of the National Academy of Sciences of the United States of America》1971,68(3):645-652
Evidence is presented to show the presence in normal rat serum of four different serum factors essential for growth of 3T3 or SV40-transformed 3T3 mouse fibroblasts: a factor that specifically promotes growth of normal 3T3 cells; two factors that specifically promote growth of transformed 3T3 cells; and a factor that sustains viability of both normal and transformed 3T3 cells in serum-free medium, probably without inducing growth of the cells. These factors are separated and partially purified. 相似文献
8.
Dieter Paul Allan Lipton Ingrid Klinger 《Proceedings of the National Academy of Sciences of the United States of America》1971,68(3):645-648
Evidence is presented to show the presence in normal rat serum of four different serum factors essential for growth of 3T3 or SV40-transformed 3T3 mouse fibroblasts: a factor that specifically promotes growth of normal 3T3 cells; two factors that specifically promote growth of transformed 3T3 cells; and a factor that sustains viability of both normal and transformed 3T3 cells in serum-free medium, probably without inducing growth of the cells. These factors are separated and partially purified. 相似文献
9.
Temperature-Dependent Properties of Cells Transformed by a Thermosensitive Mutant of Polyoma Virus 总被引:7,自引:6,他引:7
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Renato Dulbecco Walter Eckhart 《Proceedings of the National Academy of Sciences of the United States of America》1970,67(4):1775-1781
In cells transformed by ts-3, a thermosensitive mutant of polyoma virus, the loss of inhibition of DNA synthesis by topographical factors (topo-inhibition), is rendered temperature-dependent, providing evidence that the viral genome controls this essential aspect of transformation. The expression of two other attributes of transformed cells, growth in agar and serum requirement for initiation of DNA synthesis in a wound in culture, is not made temperature-dependent. In productive infection of BALB-3T3 cells by ts-3, virus-induced stimulation of cellular DNA synthesis and movement is rendered temperature-dependent. 相似文献
10.
Platelet-derived growth factor stimulates formation of active p21ras.GTP complex in Swiss mouse 3T3 cells. 总被引:39,自引:7,他引:39
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T Satoh M Endo M Nakafuku S Nakamura Y Kaziro 《Proceedings of the National Academy of Sciences of the United States of America》1990,87(15):5993-5997
The ras gene product (p21) is a GTP-binding protein and is thought to play an important role in signal transduction of growth and differentiation in many types of mammalian cells. The p21.GTP complex is an active conformation, as described previously for polypeptide chain elongation factors (EF-Tu and EF-G) and heterotrimeric GTP-binding proteins (G proteins). In the study reported here, we measured the amounts of p21-bound guanine nucleotides under various conditions in the G54 cell line, a derivative of Swiss 3T3 cells that overexpresses normal c-Ha-ras. More p21.GTP complexes were present in growing cells than in quiescent cells. When quiescent cells were stimulated with fetal bovine serum to promote DNA synthesis, p21.GTP increased approximately 2-fold. Among a number of purified growth factors, platelet-derived growth factor enhanced the formation of p21.GTP, whereas the combination of bombesin and insulin, which also induces DNA synthesis, did not. These results strongly suggest that p21 is a transducer of the growth signal from the platelet-derived growth factor receptor in Swiss 3T3 cells and that the signal is transmitted through a p21.GTP complex. 相似文献
11.
Robert W. Holley Josephine A. Kiernan 《Proceedings of the National Academy of Sciences of the United States of America》1974,71(7):2908-2911
The initiation of DNA synthesis in 3T3 cells can be controlled by a variety of different factors. Cells that are quiescent because of limited serum initiate DNA synthesis in response to the fibroblast growth factor of Gospodarowicz, to insulin, and to dexamethasone, as well as to other, unidentified biological materials. Interactions are seen between the effects of different factors. 相似文献
12.
Antitubulin agents enhance the stimulation of DNA synthesis by polypeptide growth factors in 3T3 mouse fibroblasts. 总被引:14,自引:2,他引:14
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M Friedkin A Legg E Rozengurt 《Proceedings of the National Academy of Sciences of the United States of America》1979,76(8):3909-3912
Colchicine and other antitubulin agents markedly enhanced the stimulation of DNA synthesis by combinations of various growth factors such as epidermal growth factor, insulin, fibroblast-derived growth factor, and vasopressin in serum-free cultures of several quiescent 3T3 mouse fibroblast cell lines. Enhancing effects were observed based on continuous incorporation of [3H]thymidine into DNA as well as by autoradiographic labeling of cell nuclei. The concentration of colchicine and podophyllotoxin required to produce half-maximal enhancement of DNA synthesis stimulated by epidermal growth factor and insulin was 25-50 nM. Lumicolchicine did not produce enhancing effects. The disassembly of microtubules resulting from the action of colchicine, Colcemid, and vinblastine did not inhibit the stimulation of DNA synthesis in quiescent Swiss 3T3 fibroblasts by fetal bovine serum. We conclude that the cytoplasmic microtubule network in 3T3 mouse fibroblasts does not exert a positive regulatory function in the initiation of DNA synthesis but rather can produce a constraint on the initial action of the peptide growth factors in serum-free media. 相似文献
13.
T Jippo-Kanemoto S Adachi Y Ebi H Matsuda T Kasugai S Nishikawa Y Kitamura 《Blood》1992,80(8):1933-1939
Proliferation of murine mast cells is induced by both T-cell-derived and fibroblast-derived growth factors. Because the most potent T-cell-derived mast cell growth factor, interleukin-3, promotes the migration of mast cells, we investigated whether fibroblast-derived growth factors had the chemoattractive activity as well. Conditioned medium (CM) of BALB/3T3 fibroblasts induced the migration of cultured mast cells (CMC) derived from normal (+/+) mice. BALB/3T3-CM contained the mast cell growth factor (MGF)/stem cell factor (SCF)/kit ligand (KL), which is the ligand for the receptor encoded by the W (c-kit) gene. CMC derived from the spleen of W/W mice lack the extracellular domain of the W (c-kit) receptor, and W/W CMC did not proliferate in response to BALB/3T3-CM. However, W/W CMC did migrate normally toward BALB/3T3-CM and, moreover, the antibody to the extracellular domain of the W (c-kit) receptor did not inhibit the chemoattractive activity of +/+ CMC toward BALB/3T3-CM. These results indicated that MGF/SCF/KL itself did not represent the major chemoattractive activity. On the other hand, BALB/3T3-CM induced neither proliferation nor migration of CMC derived from mi/mi mice. Both W/W and mi/mi mice are deficient in mast cells, but the present results suggest that the mechanism of the abnormality is different between W/W and mi/mi mice. 相似文献
14.
Effect of a Fibroblast Growth Factor, Insulin, Dexamethasone, and Serum on the Morphology of BALB/c 3T3 Cells 总被引:4,自引:0,他引:4
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Denis Gospodarowicz John Moran 《Proceedings of the National Academy of Sciences of the United States of America》1974,71(12):4648-4652
The effects of serum, fibroblast growth factor, dexamethasone, and insulin on the morphology of two lines of BALB/c 3T3 cells are described and illustrated. Fibroblast growth factor, a polypeptide purified from bovine brain and pituitary glands, stimulates DNA synthesis and cell division in both sparse and confluent cultures of quiescent 3T3 cells. When cells are grown in the presence of the factor, they go through one or two cycles of division and their morphology differs from that of cells either growing in 10% serum or maintained in a quiescent state in low serum; they appear rounded with long processes and show no sign of contact inhibition. When dexamethasone is present with the growth factor, the cells grow to a high density and are not contact-inhibited; they have the appearance of transformed cells. Addition of insulin or dibutyryl cyclic AMP to the cells results in a morphology similar to that of cells growing in the presence of serum. Addition of growth factor or growth factor plus dexamethasone to cells that have reached confluency in 10% serum and that are clearly contact-inhibited results in the resumption of growth. The final cell density reached is similar to that of cells growing in the presence of 30% serum with daily fluid changes. No contact inhibition is observed in either of these situations. 相似文献
15.
When skin is wounded, human keratinocytes at the wound edge stop differentiating and start migrating to resurface the wound. How this change takes place is unclear. Because keratinocytes at the wound edge are for the first time surrounded by serum rather than plasma, serum could contain some migration-promoting factor or factors that is absent in plasma. We did standard computer-assisted in-vitro migration assays of human keratinocytes in the presence of either human plasma or serum. We also did a semiquantitative western blot analysis to determine if p38 mitogen-activated protein kinase (p38MAPK) was activated by either serum or plasma. Our results showed that keratinocytes migrating on collagen in the presence of serum produced migration indices in the range of 28, whereas those in the presence of plasma were about 12--the same level as control assays without either serum or plasma. We also showed that induced keratinocyte polarisation, activation of p38MAPK, and production of matrix metalloprotease 9 are possible mechanisms for promotion of re-epithelialisation of skin wounds by human serum. 相似文献
16.
Coordinate control of 3T3 cell proliferation by platelet-derived growth factor and plasma components. 总被引:29,自引:15,他引:29
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A Vogel E Raines B Kariya M J Rivest R Ross 《Proceedings of the National Academy of Sciences of the United States of America》1978,75(6):2810-2814
DNA synthesis and cell division were measured in Swiss mouse 3T3 cells cultured in different concentrations of cell-free plasma-derived serum and increasing amounts of a platelet-derived growth factor. In plasma-derived serum alone, the cells were quiescent and they were arrested in the Go/G1 phase of the cell cycle. Addition of a platelet-derived growth factor to quiescent cells maintained in plasma-derived serum stimulated both DNA synthesis and cell division. When plasma components were present at high concentration (5%, vol/vol), the amount of platelet factor added to the cultures determined the number of cell doublings. Plasma-derived molecules were required for the platelet factor to stimulate DNA synthesis and cell division in the maximal number of cells. In addition, plasma components had to be present for recently divided cells to respond to the platelet factor. When 3T3 cells were cultured in excess platelet factor and limiting amounts of plasma-derived serum (0.5%, vol/vol), the cells underwent one doubling and then ceased to proliferate. Addition of fresh plasma-derived serum to these cells induced a second round of cell division. Plasma components and the platelet-derived growth factor acted in a coordinate fashion to regulate the proliferation of Swiss 3T3 cells. 相似文献
17.
Basic fibroblast growth factor released by single, isolated cells stimulates their migration in an autocrine manner. 总被引:18,自引:0,他引:18
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P Mignatti T Morimoto D B Rifkin 《Proceedings of the National Academy of Sciences of the United States of America》1991,88(24):11007-11011
Basic fibroblast growth factor (bFGF), a protein with angiogenic, mitogenic, and chemotactic properties, lacks a signal sequence and is not secreted via the classical secretory pathway. However, the growth factor is known to act extracellularly. Since no defined mechanism for bFGF release has been described, it has been suggested that this growth factor is released from dead or damaged cells. To test this hypothesis we characterized the effect of exogenously added bFGF and neutralizing antibody on the migration of single, isolated NIH 3T3 cells transfected with bFGF cDNA. Under these conditions the observed cell cannot be affected by bFGF derived from other cells. Cells were seeded onto colloidal gold-coated coverslips at a density of one cell per coverslip. A cell migrating on this substrate produces a track free of refringent gold particles that is measured by an image analyzer. The results showed that cell motility directly correlated with the amount of bFGF released from the migrating cells. Affinity-purified anti-bFGF antibody, but not irrelevant IgG, reduced the level of migration of the bFGF transfectants to that of the control cells transfected with the vector alone, showing that bFGF stimulates migration of the cell that releases it. Thus, bFGF is secreted by viable cells and mediates cell functions via a "true" autocrine mechanism. 相似文献
18.
E Rozengurt A Legg G Strang N Courtenay-Luck 《Proceedings of the National Academy of Sciences of the United States of America》1981,78(7):4392-4396
Addition of cholera toxin (100 ng/ml) to quiescent cultures of Swiss 3T3 cells acts synergistically with serum (2-4%), insulin, phorbol esters, epidermal growth factor, and fibroblast-derived growth factor to stimulate DNA synthesis. In the presence of insulin, cholera toxin caused a dose-dependent increase in cumulative [3H]thymidine incorporation into acid-insoluble material and in the intracellular cyclic AMP (cAMP) level. The dose--response curves for the two processes were similar. Furthermore, addition of 1-methyl-3-isobutylxanthine (15--500 microM) or of 4-(3-butoxy-4-methoxybenzyl)-2-imidazolidinone (5--100 microM), both of which are potent inhibitors of cyclic nucleotide phosphodiesterase which are potent inhibitors of cyclic nucleotide phosphodiesterase activity, stimulated DNA synthesis and increased cAMP levels in Swiss 3T3 cells. These compounds strikingly potentiated the effect of cholera toxin on DNA synthesis and on cAMP levels. When quiescent Swiss 3T3 cells were exposed to cholera toxin (100 ng/ml) and insulin at 10 micrograms/ml (4- to 7-fold increase in cAMP level) or to these agents and 1-methyl-3-isobutyl xanthine at 50 microM (35-fold increase in cAMP level), DNA synthesis began after a lag of 16 hr. These results indicate that cAMP acts as a mitogenic signal for Swiss 3T3 cells and differ from the widely held view that cyclic AMP inhibits the proliferation of fibroblast cells. 相似文献
19.
Induction of DNA synthesis in BALB/c 3T3 cells by serum components: Reevaluation of the commitment process 总被引:61,自引:39,他引:61
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W. J. Pledger C. D. Stiles H. N. Antoniades C. D. Scher 《Proceedings of the National Academy of Sciences of the United States of America》1977,74(10):4481-4485
Serum contains a growth factor derived from platelets and also growth factors derived from platelet-poor plasma. Extracts of heated (100 degrees ) human platelets function synergistically with platelet-poor plasma to induce DNA synthesis in quiescent, density-inhibited BALB/c 3T3 cells. Platelet-poor plasma alone did not induce DNA synthesis. Cells exposed to platelet extracts became competent to enter the cell cycle, but the rate of entry into the S phase depended upon the concentration of platelet-poor plasma. The time required for the induction of this competent state was a function of the concentration of the platelet extract. A 2-hr exposure to 100 mug of the platelet extract at 37 degrees caused the entire cell population to become competent to enter the S phase. At 4 degrees or 25 degrees the cells did not become competent to synthesize DNA. The platelet extract-induced competent state was stable for at least 13 hr after removal of the platelet extract; however, in the absence of platelet-poor plasma, these competent cells did not progress through the cell cycle. The addition of an optimal concentration of platelet-poor plasma (5%) to these competent cells initiated cell cycle traverse with a rapid, first-order entry of cells into the S phase beginning 12 hr after addition of the plasma. The addition of a suboptimal concentration of the plasma (0.25%) did not increase the rate of cell entry into the S phase. Thus, the induction of DNA synthesis in quiescent BALB/c 3T3 cells can be resolved into at least two phases, controlled by different serum components: (i) competence, induced by the platelet-derived growth factor; and (ii) progression of competent cells into the cell cycle, mediated by factors in platelet-poor plasma. 相似文献
20.
J M Fentener van Vlissingen E J van Zoelen P J Ursem C J Wensing 《Endocrinology》1988,123(6):2868-2877
The gubernaculum testis is the connective tissue organ that causes the testis to descend. How the process of testicular descent is regulated is not fully understood. Current hypotheses postulate that a nonandrogenic fetal testicular factor controls the first phase of descent, that is characterized by growth of the gubernaculum and transabdominal migration of the testis. When gonadal extracts from fetuses with ages corresponding to the first phase of testicular descent (50, 60, and 75 days) were tested on gubernacular cells, the growth stimulatory effect of testicular extracts exceeded the effect of both ovarian extract and fetal calf serum. Gonadal extracts from 80-, 90-, and 100-day-old fetuses showed only a minor sex difference. No sex difference or age-dependent changes were detected when fetal gonadal extracts were tested on murine 3T3 cells. Polypeptide growth factors (epidermal growth factor, insulin, fibroblast growth factor, platelet-derived growth factor, and transforming growth factor-beta) were tested for growth stimulatory activity and had only minor effects on gubernaculum cells. Fetal testicular hormones (anti-Müllerian hormone, inhibin, and androgenic steroids) did not induce initiation of DNA synthesis at concentrations that are highly bioactive in typical target systems. When testicular samples were dialyzed, the high mol wt fraction (greater than 3500) had lower growth stimulatory activity in gubernaculum cells, but not 3T3 cells. Bioactivity of ovarian extracts and fetal calf serum was not diminished after dialysis. The low mol wt fraction (less than 3500) of testicular extract was distinctly stimulatory to gubernaculum cells but not 3T3 cells, and the low mol wt fraction of ovarian extracts did not stimulate growth in either cell type. It was concluded that the fetal porcine testis during the first phase of testicular descent contains low mol wt factor(s) to which gubernaculum cells and not 3T3 cells are responsive. The bioactive fraction probably contains the factor(s) that initiate testicular descent. We suggest the name descendin for this new activity. 相似文献