Production of progesterone induced by follicle-stimulating hormone by buffalo (Bubalus bubalis) granulosa cells in vitro

Rad iolahelled1 prou.ster-one(1c 6 'II pt ogestet one) ssa", obtlainfld frtoni Amet -sham Radiochemicail 'Centet1ot t lie pit epairation of IiOs-precoated plates, 48 swcel platcs 1ssali Isa ) wsetecoated Wit~h I3't)[(1 and in IIIh1atCd OVCI e1igt ,at 371 (InI aii lattmosplic't e of 5percenit clarbon dioxsidc in air'. 'Ilc he n\t clav, Miicomed jatelv beforec the ladd it ion of gt an iLosa Cells fo the cLnltn e CsWell, theC HIWOSsa removed anid the wsells \55F ct e S shCd Wsithli 2~t)l Of Set nni ftI-ee nieidinml. \fet2 hooso cutut, 111ft medium1 WIs Cat etull tetiosed m''iii echII W55 cl WithouIt diStI.tt bing( the CCellS a ncl eplalcecl wsithI 411 11plOf freCSh Set nillt'ft ee mediuml. TI het eaf ter, theC mICCli'uIII 55 as Cotmpletely retmovt cd atid treplaced esvety dlisv. 'Ilie set urrti ft-CICedmel1iti aIdded clailsV COntaitieCd tieC teSpeCCtts'ecoticeti ft a1-tO (iOf t,s1. I'C heSample[IS Of 1niCedi,mii COlICCtcd \Vls cieStoteCl aIt -21) Luntil SLIISeC(fLeCt anaI1Ssis fo pi(opestet otie. Il1 o(cestet otie lesvels ssetre estimIated lis a cilit ctt racdi0 mmton-toassay as desc tI bed Ki amboj land Pi aklash 993 lIeittt a anllId intitetassa coeficitts oIf satiation \s ct ls flai11petCetC Pt C ogttOtIC It(icOdLItciOiti dalta ssCetC e\pt essecd as poifi rooestetrone pettCl(if O1CoLtL_ meMecCli'tii at Ctet C(tt eCtIOti tot t t tl e_SIcl1~1 ual amoun1t Of nielCi uII left Ill theC sWells sh let the mecdlintii ssaS ChangedCC Otil clas of cnIltoteC. FI aCli ctl'tiiettC s astdttiS s-iSI t toecs. 'I lie data ss et lo, ti-ansfot med befote1 analsVSIS atld thetI SLufiectecd to ati-alssis oitvat ancc sswith )tLi1Canlls tInlIItIp -lerange test. I hec basal pt ogeCstet one scC,tetoti Conftro (vsalues, obtainted rotii CCells CtlIt utCedin t le albsetice of' iI ) Is theC gi a ti,u0S lo clls \\-,-s similar oti class and 2 (if culituteC Fh, I progestetr((ti secrei(itio tic eased s(i lt ifica llif\I P ilcl)() betsweeti class 2) atid 1, anId tliele sas tlieti n d1(1Cclieteioc (It clas 4. tituig II anulosa cells wsitli the cIt dcdec closes of Iiad tio cf fecct otil fit ogestet ((ticft OcluctioCti Oti0I claIs atid 2 of coiltLut c Fli2 Hossvceset (idlay' 3 the cells ft eafcIc switli IS Ii secicttcli tiiote( ('s i p 'csI tote t ollciICetift ,ttiotis (iI,11 suiida pattet-1tisa (bSiiset sec I (tiVc 4 Oif CLultute-. IlteCttilieIti ssifth esveti the lossest c oticetittat ioti oIf uII tha"t is, ti i,IIl teCSulted iti al 25 pct, Ccc'ti tid 2(t petl cetitit1clIase P It'll's-) Ilit litogeCstet otie sectretioti owisetie anioutits sect eted bs the cotitr-IS((l dtcavss3 atic 4 Of CLl1tLn C, 1-CSpeCftsVCIVS TO theC antlior-s Iktossledge. thils is lie fitlst teCpoItt1 (If h desVelOptiietit Of a1 C LiItLtteSVSstetii IOtbIioffO alo gr aiulOSa CCells 55hilch is clap.tble Of Supp)[OlttitIg hot1 tiiotie fit 0CductOin. A\ ise iti basal fitrogestet (otic sec-teion ssitli tiliii act ees ss\ell ss itli theC eSuLtS Otf ptFesOuIS StUdieS iti sshiCSCliset tii f ees svtetiis ssC'te Used fol thCelViti O Cti tLrre Oit bOVsItieganu1,1isa C('Ifs ft_II Stiiliiall ada a1tici (itliet sI999), large' Roillic IC ticl other-s 1996, 1998 11 aiclfat(' ge preosLu latot V BlIernd tSti aic1d othlets 1995) follicles, atlicl of usI'I iti1giatiu.losa celS froitii Smiall atcllag f'ollicles C amipbell atin tit 19 oails CCellS utIidcltO gocifferenltiatioti atid Ltlueitsatt(inl s\ IeiC1 tlIesV ateI' Culturied Iti svittro, leaditig to ali tict ease ill pt dioestet one sectti-citio. 1 lie tiiagii it ude atic tate of tlic tease il proge('Stet mie sectectioiti dcefieiclpitia PI11ifs1 up_OnI ieleCee(1(t if cl feCreltitatioti (if the cells sse liticlturitigM ieg(iIis. Ci-atiull-oSa' FIG 1: Progesterone secretion during four days of culture of buffalo granulosa cells isolated from small follicles and cultured in serum-free medium without the addition of bovine folliclestimulating hormone. Different letters indicate significant differences (P<0-05)