(diam inobenzidine,dab ) ; 0. 5 mmo lgl, : N ac l 8 g, KC l 0. 2 g T ris 3
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1 34 8 ( ) V o l. 34 N o Jour. of N o rthw est Sci2T ech U niv. of A gri. and Fo r. (N aṫ Sci. Ed. ) A ug Ξ 1, 2, 1 (1, ; 2, ) [ ] g,,, 85%,,, 8 ; g; Fas2L, 95% ;, [ ] ; ; [ ] Q [ ] A [ ] (2006) ,, [1 ],,,,, [224 ],,,,, Feu lgen,,, Feu lgen, g,,, W istar, M edium 199 g Sigm a ; Fas2L SABC ; 3, 32 (diam inobenzidine,dab ) ; 0. 5 mmo lgl, : N ac l 8 g, KC l 0. 2 g T ris 3 g ml, ph 7. 4, V ( ) V ( ) = W istar, 75%,,,,,, 4 H ankπs,,, rgm in 5 m in, 1 m ggml, m in, mm, H ankπs rgm in 5 m in, 1 ml 0. 5 mmo lgl, 1 m in, rgm in 5 m in,, 10% M 199, T 75, 37 5% CO 2 7 h, Ξ [ ] [ ] 863 (2003AA ) [ ] (1981- ),,,, [ ] (1956- ),,,,,
2 40 ( ) h, 3 ml 0. 5 mmo lgl 3 m in,, mmo lgl, H ankπs gml,, ;, H E,, H E cm 2,, gml 96, 200 ΛL 10, 48 h 1, 10, 12,, 5 m ggml M T T 100 ΛL, 37 4 h,, 150 ΛL, 10 m in, 490 nm (A ),, A 1. 5, 95% 10 m in, (Pho sphate Buffer So lu2 tion, PBS) 2 Fas2 L, 0. 1% T ri2 ton X % H 2O 2 PBS 10 m in, PBS 2, 5% 10 m in, Fas2L, 37, IgG, m in,, m in, DAB,,, PBS, Fas2L 10, 4, ,,,, ,, ( 1), 85%, 1 (100 ) F ig. 1 Serto li cells excluding T ypan blue (100 ) H E, 3 h,,,, 7 8 h,, ( 2 A ) 2 d,,,,, :, ( 2 B ) ;, 3 d,, 50 :,,, ( 2 C) ; 100 :, ( 2 D ) 5 d, H E,,,, ( 2 E)
![Serto li cells cultured fo r 3 d (100 ) ; E. Serto li cells of H E staining (100 ) 2. 2. 2,,,,, ( 3), [2 ] 2. 2. 3 4,, A, 8, 4 F ig.](/docs-images/92/110959399/images/3-2.jpg "4 Grow th graph of rats Serto li cells 3 (20 000 ). F ig. 3 U ltrastructure of nuclei in Serto li cells of rats (20 000 ).")
3 8 : 41 2 A. 7 h (100 ) ; B. 2 d (400 ) ; C. 3 d (50 ) ; D. 3 d (100 ) ; E. H E (100 ) F ig. 2 R ats Serto li cells of different culture tim es A. Serto li cells cultured fo r 7 h (100 ) ; B. Serto li cells cultured fo r 2 d (400 ) ; C. Serto li cells cultured fo r 3 d (50 ) ; D. Serto li cells cultured fo r 3 d (100 ) ; E. Serto li cells of H E staining (100 ) ,,,,, ( 3), [2 ] ,, A, 8, 4 F ig. 4 Grow th graph of rats Serto li cells 3 ( ). F ig. 3 U ltrastructure of nuclei in Serto li cells of rats ( ). Satellite nucleo som es around the nucleo lus 2. 3 Fas2L Fas2L, ( 5), Fas2L,, 95%
4 42 ( ) 34 5 Fas2L (200 ) F ig. 5 R at Serto li cells of Fas2L imm uno2 cytochem ical staining (200 ) , Greene [1 ],,, 1993, Selaw ry [5 ],,, Χ2 (Glu tam yl T ran spep tidase, Χ2GT P) 1 2 (SGP, SGP21, SGF22) 21 ( In su lin2like facto r, IGF21) 2Α, [324, 6 ],,,,,, [7 ],,,,, Fas TN F, Fas2L, Fas T B, Fas2L, Fas T, [8 ] Fas2L,, Fas2L, Fas2L, Fas2L Fas2L [9 ], Fas2L Fas,, 1, Fas2L,,, [ ] [ 1 ] Jannette M D, Ray V R, Grego ry S K, et al. H arnessing the imm unomodulato ry p roperties of serto li cells to ennable xeno transp lantation in type g diabetes[j ]. Imm uno l Invest, 2003, 32 (4) : [ 2 ] Ko rbutt G S, E llio tt J F, Rajo tte R V. Co transp lantation of allogeneic islets w ith allogeneic testicular cell aggregates allow long2term graft survival w ithout system ic imm uno supp ression[j ]. D iabetes, 1997, 46: [ 3 ] O thberg A I,W illing A E, Cam eron D F, et al. T roph ic effect of po rcine serto li cells on rat and hum an ventral m esencephalic cells and hn T neurons in v itro[j ]. Cell T ransp lant, 1998, 7 (2) : [ 4 ],. Serto li [J ]., 2003, 24 (3) : [ 5 ] Selaw ry H P, Cam eron D F. Serto li cell2enriched fractions in successful islet cell transp lantation [J ]. Cell T ransp lant, 1993, 2: [ 6 ] L uca G, Calvitti M, N eri L M, et al. M itogenic effects of rat serto li cells on adult homo logous islet beta2cells: in v itro and in v ivo studies[j ]. T ransp lant P roc, 2001, 33 (1g2) :
5 8 : 43 [ 7 ] Cam eron D F,M uffly K E. Ho rmonal regulation of sperm atid binding[j ]. Cell Sci, 1991, 100 (30) : [ 8 ] Griffith T S, B runner T, F letcher S M. Fas ligand2induced apop to sis as a m echanism of imm une p rivilege[j ]. Science, 1995, 270 (5239) : [ 9 ] Bellgrau D, Go ld D, Selaw ry H, et al. A ro le fo r CD 95 ligand in p reventing graft rejection[j ]. N ature, 1995, 377: Iso lation, purification and iden tification of testis Serto li cells in p repubertal rats KONG Qing-xue 1, 2, ZHANG Y ong 1 (1 Colleg e of A nim al S cience,n orthw est A & F U niversity, Y ang ling, S haanx i , Ch ina; 2 T issue E ng ineering R esearch Center,A cad emy of M ilitary M ed ical S cience, B eij ing , Ch ina) Abstract: R ats Serto li cells w ere iso lated, purified and iden tified by one2tim e digestion w ith co llagenase V, differen tial adhesion, low o smo tic buffer treatm en t and imm unoh istochem ical stain ing. R esu lts demon2 strated that th is iso lation and purification m ethod of Serto li cells did little harm to the cells. T he p rim arily iso lated Serto li cells had good cell viab ility; cell su rvival rate reached 85% and there w as no b ig cell co rp s; P lan ted in flask s, cell p ro liferated rap idly and reached the p ro liferation peak at day 8 po stcu ltu red. Cell yield w as cell g; M o st acqu ired cells w ere Fas2L po sitively stained, and cell purity cou ld reach 95% ; the nucleo lu s of the cells w as clear, and there w ere satellite nucleo som es around the nucleo lu s. Key words: p repubertal rat; Serto li cell; iso lation purification ( 38 ) Abstract ID: (2006) EA Exp ression of IFN 2Χ imm uno reactivity on H PG ax is of dairy goat du ring early p regnancy ZHAO Hui-y ing, SUN J ian-hong, XU Y ong-p ing, CHEN Shu- lin, M A Y e-fe i,l IY i- shu,l IU Gen- sheng,longm in (Colleg e of A nim al S cience and T echnology,n orthw est A & F U niversity, Y ang ling, S hanx i , Ch ina) Abstract: To p robe in to the imm unomodu lation of IFN 2Χ to hypo thalam ic2p itu itary2gonadal ax is, the distribu tion of IFN 2Χ imm uno reactivity on H PG ax is of dairy goat du ring early p regnancy w as determ ined by SP m ethod of imm uno2h istochem istry. T he resu lt show ed that IFN 2Χw as distribu ted over 20 nucleu s of the hypo thalam u s, such as nucleu s m am illaris m edialis, nucleu s p re2op ticu s m edialis, area an terio r hypo tha2 lam i, nucleu s ven trom edialis, nucleu s reun ion s, nucleu s paraven tricu laris, nucleu s sup ra2op tic, and etc. M any of IFN 2Χ po sitive nerve fibers w ere found in sup ra2op tic nucleu s, paraven tricu lar nucleu s, nucleu s m am illaris m edialis and etc. N erve fibers and astrocytes IFN 2Χ imm uno reactive po sitive cells sp readed in periven tricu lar nucleu s, nucleu s p re2op ticu s lateralis, nucleu s p re2op ticu s m edialis and etc. A t the sam e tim e, IFN 2Χ is also ob served obviou sly in adenohypophysis and neu rohypophysis. IFN 2Χ also exp resses in fo llicu lar and co rpus lu teum. T he resu lts suggest that IFN 2Χ is related to the activity of neu roendocrine of an im als, and is concerned w ith the rep roductive imm un ity regu lation in imm uno2neu ro2endocrine netw o rk. Key words: IFN 2Χ; H PG ax is; early p regnancy; dairy goat; SP m ethod
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