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1.
The percentage of sheep embryos that continued to develop after collection and immediate transfer on d 2 after estrus was similar when phosphate-buffered saline with 10% fetal calf serum (PBSFCS, 45%), physiological saline (50%), or tissue culture medium 199 supplemented with 10% fetal calf serum (M199FCS, 47%) was used to flush embryos from oviducts. Co-culture of sheep embryos for 3 d with oviductal cells tended (P = .1) to reduce the percentage of embryos that developed to fetuses after transfer compared with those embryos transferred immediately. Tissue culture medium 199 supplemented with .3% BSA (M199BSA) was an adequate substitute for M199FCS for culture of sheep oviductal cells if tissue culture wells were pretreated with fibronectin. Estradiol in concentrations from 10 to 1,000 pg/ml and progesterone at concentrations of 1 or 10 ng/ml in M199BSA failed to stimulate embryo development during 3 d of co-culture beyond that seen in co-culture with M199FCS or M199BSA without added steroid. Transfer of sheep embyros co-cultured for 3 d in M199BSA or M199FCS to recipients synchronized with donors resulted in about 19% of the embryos developing to fetuses, whereas transfer to recipients that were in estrus 24 h after donors resulted in 33% of embryos developing to fetuses. The significant (P less than .05) improvement for delayed recipients may reflect the relatively lesser developmental rate of co-cultured embryos compared with that of embryos in vivo. Embryo development into fetuses was similar after co-culture in M199FCS or M199BSA co-cultures; therefore, serum is not required for the co-culture of sheep embryos.  相似文献   

2.
The effects of different cell monolayers on in vitro development of early bovine embryos derived from in vitro maturation and fertilization were examined in this study. Early embryos (four to eight cells) were randomly allocated to bovine granulosa cell (GC), oviductal cell (OC), or uterine cell (UC) monolayers in Exp. 1 and to GC, skin cell (SC; from 10-d-old chicken embryos), testicular cell (TC; from 10-d-old mouse), and liver cell (LC; from 10-d-old chicken embryos) monolayers in Exp. 2, and cultured for 6 d at 38.6 degrees C in a humidified atmosphere of 5% CO2 in air. The culture medium was 12.5 mM HEPES TCM 199 supplemented with 1% calf serum and 1 mM sodium pyruvate. In Exp. 1, the percentage of four- to eight-cell embryos that developed to blastocysts on GC, OC, and UC monolayers was 26.9 (28/104), 37.5 (39/104), and 39.2 (40/102), respectively. In Exp. 2, the percentage of four- to eight-cell embryos that developed to blastocysts on GC, SC, TC, and LC monolayers was 53.3 (40/75), 42.9 (33/77), 49.3 (37/75), and 44.3 (35/79), respectively. There were no significant differences in development among groups in either experiment.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

3.
Development competence and pregnancy rate of in vitro-produced (IVP) dromedary embryos were studied in two culture systems: (i) semi-defined modified medium (mKSOMaa) and (ii) co-culture using camel epithelial oviducal cells. Five hundred and three cumulus-oocytes complexes (COCs) were selected, allowed to mature, fertilized and cultured in vitro (38.5 degrees C; 5% CO2, maximum humidity > 95%, with concentration of oxygen of 5% for semi-defined medium and 20% for co-culture cells). Maturation was accomplished by incubation in TCM-199 medium supplemented with 10% heat-treated foetal calf serum (FCS), 10 ng/ml epidermal growth factor, 1 microg/ml follicle-stimulating hormone, 1 microg/ml oestradiol and 500 microM cysteamine for 30 h. In vitro fertilization (IVF) was performed using fresh semen (0.5 x 10(6) spermatozoa/ml in modified TALP solution). Fertilized COCs were denuded by vortexing, then cultured in either mKSOMaa (10% heat-treated FCS was added 24 h post-IVF), under 5% O2 and 90% N2 (group 1; n = 249) or with dromedary epithelial oviducal cell monolayers in TCM-199 with 10% heat-treated FCS under 20% O2 (group 2; n = 254). The rate of cleavage was significant higher (p < 0.05) for group 1 (63%, 156/249) than for group 2 (51%, 130/254). No significant difference was found between the two groups in the rate of development to blastocyst (21% vs 16.5%) and their hatchability (21% vs 14%). Pregnancy rates were similar for the first 60 days. However, all pregnancies were lost after 60 days with the exception of two of six (33%) from recipients of hatched blastocysts from group 1. We conclude that both systems support in vitro production of dromedary embryos by in vitro maturation (IVM)/IVF of oocytes. However, embryos obtained by culture in the semi-defined medium (mKSOMaa) appear to have a better in vivo development ability.  相似文献   

4.
Two experiments were conducted to examine the temporal aspects of luteal resistance to the luteolytic effect of prostaglandin (PG) F2 alpha during early pregnancy. In Exp. 1, 14 pregnant and 12 nonpregnant ewes were treated with PGF2 alpha either on d 10 or 13 post-estrus. Jugular venous blood samples were collected at -30 min, 0, 6, 12, 18, 24, 30 and 36 h post-injection for quantification of progesterone. The difference (delta P) between pre-treatment and post-treatment concentrations of progesterone was calculated for each ewe. There was a significant interaction between pregnancy status and day of treatment on delta P (P less than .05). Pregnant and nonpregnant ewes treated on d 10 showed a large delta P. A large delta P also was observed in nonpregnant ewes treated on d 13 post-estrus. However, delta P in pregnant ewes treated on d 13 was smaller than in the other three groups (P less than .05). The temporal patterns of concentrations of progesterone in serum were different among treatment groups (P less than .05). A suppression in the concentration of progesterone was observed by 24 h post-injection in all four treatment groups. Progesterone returned to pre-treatment levels only in pregnant ewes treated on d 13. In Exp. 2, 47 pregnant ewes were treated with PGF2 alpha on d 10, 13, 16, 19, 22, 26 or 30 postestrus. Blood samples were collected and data were analyzed as described for Exp. 1.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

5.
A surgical uterine flush technique was evaluated for percentage of ova/corpora lutea (CL) collected and development of reproductive adhesions in 79 superovulated ewes. The median collection rate for ova/CL was 70%, and 47 of 79 ewes had greater than or equal to 67% ova/CL. At 50 to 80 days after uterine flushing, celiotomies were performed on 50 of the 79 ewes to evaluate the reproductive tract for adhesions. Adhesions of the reproductive tract were not found. Thirty ewes were given prostaglandin F2 alpha, were mated, and became pregnant. The ova collection rates were comparable with rates reported in ewes in which oviductal flush methods were used; however, adhesions did not develop and reproductive function was maintained after surgery.  相似文献   

6.
按照多房棘球绦虫幼虫-泡球蚴培养的培养基（RPMI-1640、M199和MEM）分为3组：Ⅰ组为含10％胎牛血清的RPMI-1640;Ⅱ组为含10％胎牛血清的MEM;HI组为含10％胎牛血清的M199。将泡球蚴在3种细胞培养液中进行培养,观察其存活、生长以及发育情况。结果显示,培养9d的泡球蚴的成活率分别为：Ⅰ组90．10％、Ⅱ组50．25％、Ⅲ组22．03％;成囊率分别为：Ⅰ组57．12％、Ⅱ组63．15％、Ⅲ组48．17％;头节外翻率分别为：Ⅰ组98．28％、Ⅱ组88．65％、Ⅲ组75．50％。可见,大多数虫体在早期向囊发育,一部分虫体头节外翻,并伴有规律的伸缩运动,但随时间的延长虫体运动减缓,又向囊蚴发育。通过对多房棘球绦虫泡球蚴的体外培养,初步表明合有10％小牛血清的细胞培养基RPMI-1640较适合泡球蚴的生长发育,为研究寄生虫发育提供了最基本的数据资料。  相似文献   

7.
屠宰绵羊卵巢卵母细胞的体外培养   总被引：2，自引：0，他引：2  
为了使屠宰绵羊卵巢卵母细胞能够用于体外受精,本文着重探索了使卵巢卵母细胞体外培养成熟的方法和条件。实验结果表明,以TCM-199加10％FCS作为基本培养基,培养24～25小时,可以使绵羊卵巢卵母细胞培养成熟,其成熟率可达55.5％(435/784)。如果在培养液内添加hCG(0.02mg/ml)或LH(0.01mg/ml),并且尽可能保持卵丘细胞的完整,则可以使成熟率提高到76.9％(140/182)～82.9％(112/135)。  相似文献   

8.
研究不同培养体系对胎牛成纤维细胞体外培养的影响及用牛血清白蛋白代替血清培养胎牛成纤维细胞的可行性。利用M199、DMEM、α-MEM、DMEM/F124种培养体系通过组织块贴壁培养对成纤维细胞体外培养液进行筛选,以α-MEM组细胞生长状况较好。分别用含2、4、6、8、10mg/mL BSA的α-MEM培养液对胎牛成纤维细胞进行原代及传代培养,5种浓度的BSA对原代培养时细胞开始游离出组织块的时间影响不明显,均在培养后的48h有成纤维细胞和上皮细胞混合游离出,但在传代培养时,胎牛成纤维细胞在8mg/mL BSA浓度的α-MEM中贴壁率较高。结果表明:培养胎牛成纤维细胞时,可用BSA代替血清,较适宜的培养体系为含8mg/mL BSA的α-MEM培养液。  相似文献   

9.
Ewes were treated with a medroxyprogesterone acetate (MAP) sponge for 8 d followed, at sponge removal, with 500 IU pregnant mare serum gonadotropin (PMSG) at d 30, 40 or 50 (d 0 = lambing) to induce estrus. Dry and lactating ewes were divided into equal numbers at each postpartum day and bred at estrus. Conception rates and number of accessory sperm were determined by flushing the oviducts 3 d after mating and examining the recovered ova. There was no effect (P greater than .05) of lactational status on conception rates. Conception rates increased (P less than .05) from d 30 (10%) to d 40 (45%) and from d 40 to d 50 (80%). There were fewer (P less than .05) ova with accessory sperm (5/26) in d-30 ewes compared with d-40 (10/27) or d-50 (12/24) ewes. In Exp. 2, ewes were assigned to two groups after receiving PMSG on d 30: 1) mated naturally or 2) inseminated during laparotomy near the uterotubal junction (UTJ). Dry and lactating ewes were divided evenly within each of the two treatments. Oviducts were flushed and ova were examined for cleavage. The conception rate was 60% in ewes that were inseminated in the UTJ vs 10% in ewes mated to rams (P less than .05). Lactational status had no effect on results. In conclusion, conception rates in postpartum ewes treated with MAP sponge and PMSG increased from postpartum d 30 to d 50 with natural breeding, and d-30 conception rates were increased over natural mating by insemination into the uterine horn near the UTJ.  相似文献   

10.
Three studies were conducted to investigate concentrations of estrogen and progestin receptors in ewe myometrium during gestation. In the first two studies, concentrations were found to be similar in pregnant and nonpregnant ewes on d 5, 9, 12 and 14 postestrus. The receptor concentrations were relatively high on d 5 and were lowest on d 14. Apparently concentrations of myometrial steroid receptors during the luteal phase are commensurate with the successful establishment of pregnancy. The concentration of both types of receptors increased from d 14 to 35 in pregnant ewes. This change was associated with a concomitant increase in the amount of DNA and protein in cells. In Exp. 3, it was found that the concentrations of the estrogen receptor remained fairly constant after d 45 until late gestation, while the concentration of cytoplasmic progestin receptor increased after d 65. The ratio of RNA to DNA appeared to increase throughout pregnancy. Progestin receptor increases may be involved in the continued growth and quiescence of the uterine myometrium.  相似文献   

11.
The present experiment was designed to identify possible effects of sera and steroid hormones added to a co-culture with bovine oviduct epithelial cells on embryonic development in vitro. Bovine oocytes were matured in vitro for 24 h and then fertilized in vitro using swim-up and heparin-treated, frozen-thawed spermatozoa. At 18 and 20 h after insemination, oocytes were cultured for 3 or 7 d in a co-culture system with bovine oviduct epithelial cells containing either fetal calf serum (FCS) or estrous cow serum (ECS) and one of six hormonal additions (none, 1 or 10 micrograms/ml estradiol [E]; 1 microgram/ml progesterone [P]; 1 microgram/ml E + P; and 10 micrograms/ml E + P). A total of 2,666 oocytes were cultured for 3 d and examined for cleavage. Of those, 2,280 oocytes were cultured up to 7 d for development to the late morula or blastocyst stage. Greatest cleavage rates for 2- to 8-cell and 8-cell stages were observed in FCS (71 and 24%) and ECS (66 and 23%) without steroid addition. For development into blastocysts, no serum effect was observed. Greatest rates for development into blastocysts were observed in FCS (14%) and ECS (16%) without steroid addition. These results indicate that addition of E and P at the doses and combinations tested did not enhance developmental capacity of in vitro fertilized bovine oocytes. Compared with FCS, ECS tended to increase cleavage rates and development into blastocysts.  相似文献   

12.
Immunosuppressive activity of conditioned medium from cultured ovine, caprine, and hybrid trophoblast tissue was examined. Conceptuses were obtained from naturally mated donor ewes and does at d 20 of gestation and trophoblast tissue was cultured for 24 h in medium supplemented with 15% calf serum and 1% antibiotic/antimycotic. Conditioned medium was added to pokeweed mitogen-stimulated sheep and goat lymphocyte cultures. Quantification of [3H]thymidine uptake by cells was used to measure lymphocyte proliferation. Ovine, caprine, and hybrid conditioned medium effectively suppressed sheep and goat lymphocyte proliferation (P less than .01). There were no differences (P greater than .05) between the immunosuppressive activity of the three tissue types on either sheep or goat lymphocytes. For all treatment groups, sheep lymphocytes were suppressed more than goat lymphocytes (P less than .05). These results indicate that, at d 20 of gestation, sheep x goat hybrid trophoblast tissue is capable of suppressing pokeweed mitogen-stimulated lymphocyte proliferation.  相似文献   

13.
The aim of the present study was to address the effect of resveratrol-mediated upregulation of sirtuin 1 (SIRT1) during oocyte maturation on mitochondrial function, the developmental ability of oocytes and on mechanisms responsible for blockage of polyspermic fertilization. Oocytes collected from slaughterhouse-derived ovaries were cultured in TCM-199 medium supplemented with 10% FCS and 0 or 20 µM resveratrol (Res). We examined the effect of Res on SIRT1 expression in in vitro-matured oocytes (Exp 1); fertilization and developmental ability (Exp 2); mitochondrial DNA copy number (Mt number), ATP content and mitochondrial membrane potential in matured oocytes (Exp 3); and the time required for proteinase to dissolve the zona pellucida following in vitro fertilization (as a marker of zona pellucida hardening), as well as on the distribution of cortical granules before and after fertilization (Exp 4). In Exp 1, the 20 µM Res treatment upregulated protein expression of SIRT1 in oocytes. In Exp 2, Res treatment improved the ratio of normal fertilization and the total cell number of blastocysts. In Exp 3, Res treatment significantly increased the ATP content in matured oocytes. Additionally, Res increased the overall Mt number and mitochondrial membrane potential, but the effect was donor-dependent. In Exp 4, Res-induced zona hardening improved the distribution and exocytosis of cortical granules after in vitro fertilization. In conclusion, Res improved the quality of oocytes by improving mitochondrial quantity and quality. In addition, Res added to the maturation medium enhanced SIRT1 protein expression in oocytes and improved fertilization via reinforcement of the mechanisms responsible for blockage of polyspermic fertilization.  相似文献   

14.
Porcine embryos were flushed from mated donors and examined for cleavage stage. One- and two-cell embryos were randomly allotted to one of the five following in vitro treatments: M199 with Earle's salts, a modified Tyrode's medium (TL), TL supplemented with 10 mM N-2-hydroxyethyl-piperazine-N'-2-ethanesulfonic acid (HEPES) (TLH), TLH supplemented with 5.5 mM glucose (TLHG), or TLH supplemented with 5 mM glutamine (TLHGL). The bicarbonate concentration of TLH, TLHG, and TLHGL was 2 mM, compared with the 25 mM concentration in M199 and TL. Embryos in M199 and TL were incubated in 95% air:5% CO2 at 39 degrees C. Those in the remaining three treatments were incubated in air at 39 degrees C. Embryos incubated in TL and M199 did not develop past the four- to eight-cell stage, whereas the proportions of embryos developing to the compact morula or blastocyst stage by d 7 of culture in the other treatments were as follows: TLHG, 49.1%; TLHGL, 59.4%; TLH, 63.5% (P less than .005). These results indicate that porcine embryos can be cultured from the one-cell stage to blastocyst in a simple HEPES-buffered medium in air. The ability of porcine embryos to develop without supplemental CO2 may be an important finding for use in situations in which embryos must be transported for long periods before embryo transfer.  相似文献   

15.
Cysteine supplementation to in vitro maturation (IVM) media of bovine oocytes increases cellular glutathione production. Beneficial effects of growth factors for improving the rate of blastocyst development have been reported, but combined effects are unknown. This study was conducted to determine the additive effect of cysteine with epidermal growth factor (EGF) and/or insulin-like growth factor-I (IGF-I) on embryo development. Bovine oocytes from slaughterhouse ovaries were matured in TCM-199 (control), with or without the addition of 0.6 mm cysteine (C) at 0 or 12 h of maturation. After in vitro fertilization, embryos were allocated to culture treatments containing synthetic oviductal fluid medium. Culture treatments included fetal calf serum (FCS, 4%) alone; IGF-I (100 ng/ml); EGF (10 ng/ml); and IGF-I + EGF (100 + 10 ng/ml). Although rates for blastocysts development were not different among treatments, an increased proportion of embryos attaining morula formation was achieved when cysteine was added to the maturation media (12 h C IGF-I + EGF, 41.4%; 0 h C EGF, 40.0%) as compared to control (FCS: 34.6%). When cysteine treatments were combined, percent cleavage was greater for IGF-I + EGF (70.8%) compared to FCS (61.2%). The abundance of mRNA from the apoptotic genes, Bax and Bcl-2, and the oxidative stress genes, copper (Cu)-zinc (Zn) superoxide dismutase (SOD) and manganese (Mn) SOD in embryos was assessed. No treatment effect was observed on the expression of these genes. In conclusion, supplementation of cysteine during IVM of oocytes, in conjunction with growth factors could effectively be used as a replacement for FCS.  相似文献   

16.
Two experiments were conducted to examine the milk producing ability of Western White-Faced sheep and to identify traits that correlate well with milk production. In Exp. 1, 31 Targhee ewes were milked and five samples were taken during 107-d lactations in which the ewes nursed twin lambs. Milk yield and composition, lamb weights, ewe weights, wool growth, and udder size also were measured. In Exp. 2, 24 ewes (Rambouillet x Finn-Dorset) were separated from their lambs at 7 wk and milked twice per day for eight more weeks, during which milk yield and composition, feed consumption, udder width, and ewe weights were measured. Results from Exp. 1 showed that lamb 30-d weights, ewe weights at breeding time, and udder width at peak lactation were highly correlated with suckled milk yield (r = .81, .75 and .66, respectively). Results from Exp. 2 indicated that lamb weights and ewe weights were not useful for predicting milk yield in dairy ewes, but feed intake and udder width were (r = .74 and .86, respectively). Single-day milk yield measurements were excellent estimators of total lactation yield in both experiments. Milk yields averaged 1,714 g/d in the suckled ewes and 477 g/d in the dairy ewes.  相似文献   

17.
Contents: The present study was undertaken to examine if bovine oocytes matured either in vivo (Experiment I) or in vitro (Experiment II) and fertilized by frozen-thawed spermatozoa in vitro could produce normal calves. In experiment I two one-cell ova were transferred to the oviducts (2 × 2) of a synchronized, final recipient which ultimately became pregnant and delivered a normal bull calf after 275 days of gestation. In experiment II 54 one-cell ova were transferred to the oviduct of an intermediate recipient from which 15 blastocysts were recovered by non-surgical flushing of the uterus 6 days later. Subsequent transfer of 2 blastocysts to the final recipient by a non-surgical technique resulted in pregnancy and birth of a normal heifer calf after 281 days of gestation. The present study confirms that calves can be born from in vitro fertilization of in vivo and in vitro matured oocytes and demonstrates for the first time that calves can be born after non-surgical transfer of bovine oocytes both matured and fertilized in-vitro and incubated in cattle oviducts. Inhalt: Kälbergeburten nach In-vitro-Befruchtung von Oozyten In dieser Untersuchung wurde geprüft, ob Rinderooryten nach Reifung in vivo (Exp. 1) oder in vitro (Exp. 2) und anschlieβender In-vitro-Befruchtung sich zu normalen Käl-bern entwickeln können. Im ersten Experiment wurden zwei befruchtete, aber noch nicht geteilte Eier in den Eileiter eines synchronisierten Empfangertieres transferiert, und nach 2 75 Tagen wurde ein normales Bullenkalb geboren. Im zweiten Experiment wurden 54 in vitro gereifte und befruchtete, noch ungeteilte Eier in die Eileiter eines Zwischenempfängers übertragen und nach 6 Tagen durch unchirurgische Uterusspülung 15 Blastozysten wiedergewonnen. Ein nachfolgender unchirurgischer Transfer von 2 Blastozysten in ein Empfängertier führte zur Trächtigkeit und nach 281 Tagen zur Geburt eines normalen Kuhkalbes. Diese Ergebnisse beweisen, daβ durch In-vitro-Befruchtung von in vivo und auch in vitro gereiften Oozyten Kälbergeburten möglich sind.  相似文献   

18.
Viability of stored equine embryos   总被引：1，自引：0，他引：1  
Equine embryos were recovered nonsurgically 6.5 d after ovulation (Exp. 1) and those greater than 200 microns were stored in one of three media: 1) Ham's F10 + 10% fetal calf serum (FCS) under 5% CO2, 5% O2 and 90% N2 at 24 C (Ham's F10); 2) Minimal Essential Medium with Hank's balanced salts + 10% FCS in air (MEM) at 24 C or 3) MEM at 5 C n = 10/treatment). Embryos less than or equal to 200 micron (n = 10) were bisected microsurgically; one-half of each embryo was stored in Ham's F10 and the other half in either Dulbecco's phosphate-buffered saline + 10% FCS in air at 24 C (DPBS), or MEM in air at 24 C. At 0, 12 and 24 h, embryos were: 1) measured; 2) assigned a developmental score of 1 to 4 (1 = tight morula, 4 = expanding blastocyst) and 3) assigned a quality score of 1 to 5 (1 = excellent, 5 = degenerate). Whole embryos stored in MEM at 5 C or 24 C did not (P greater than .05) advance in development by 24 h, whereas those stored in Ham's F10 at 24 C were more (P less than .05) advanced (i.e., higher developmental score) by 24 h. From 0 to 24 h, 1 of 10, 6 of 10 and 7 of 10 whole embryos developed when stored in MEM 5 C, MEM 24 C and Ham's F10 24 C, respectively. Embryo quality was better at 24 h (P less than .05) for embryos stored in Ham's F10 at 24 C compared with MEM at 5 C.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

19.
不同培养液及血清浓度对绵羊孤雌生殖胚胎发育的影响   总被引：1，自引：0，他引：1  
体外成熟的绵羊卵母细胞，经5 μmol/L A23187 5 min和2 mmol/L 6-DMAP 4 h激活后，分别在SOFaa、M-199两种培养液中进行培养，试验比较了SOFaa、M-199液分别添加BSA和不同浓度（10%、20%）胎牛血清对绵羊孤雌生殖胚体外发育的影响。M-199+FCS组的卵裂率显著低于SOFaa+BSA和SOFaa+FCS组（P<0.05），SOFaa+BSA组的囊胚率显著低于M-199+BSA、SOFaa+FCS、M-199+FCS三组（P<0.05）；SOFaa+10% FCS与SOFaa+20% FCS组的卵裂率及囊胚率均无显著差异，M-199+10% FCS和M-199+20% FCS组的卵裂率及囊胚率间均无显著差异。结果表明：SOFaa比M-199更适合绵羊孤雌生殖胚体外发育，此外添加10%的FCS即可达到较好的培养效果。  相似文献   

20.
An E-rosetting reaction is described which gave 92.1%±2.4 (mean±S.D.) E-rosettes with bovine fetal thymocytes and 48.2%±8.4 with with bovine peripheral blood leukocyte (PBL) preparations. Both culture conditions and culture medium were critical factors in obtaining maximal and reproducible E-rosette numbers. Optimum rosette formation occurred when bovine PBL and neuraminidase treated sheep erythrocytes (nSRBC) were reacted in L-15 culture medium supplemented with 10% fetal calf serum (FCS). Other media including 100% FCS, MEM with 10% FCS, and RPMI-1640 with 10% FCS were less satisfactory. Cultural conditions found to be optimal for enumeration of bovine E-rosettes are similar to those reported as optimal for detection of human T cells. The specificity of rosette formation by bovine thymus derived (T) lymphocytes was shown by demonstration of (1) rosettes and surface membrane immunoglobulins (mIg) on different cells in PBL, (2) rosette formation by the majority of fetal thymocytes, and (3) no inhibition of rosette formation by anti-immunoglobulin serum. Using the E-rosette and mIg assays for presumptive bovine T and B lymphocytes, respectively, it was possible to differentiate from 57.5 to 90% (75.2%±9.3) of cells in bovine PBL preparations, and from 90.2 to 97.5% (94.2%±2.1) of cells in bovine fetal thymocyte preparations into T and B cells.  相似文献   

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