32 3 ( ) V o l. 32 N o. 3 2004 3 Jour. of N o rthw est Sci2T ech U niv. of A gri. and Fo r. (N aṫ Sci. Ed. ) M arch 2004 Ξ,,,, (, 712100) [ ],,,, ;,, ;,, ( );,,,,, ;, 2 4 ;, ; ; ; [ ] ; ; ; [ ] S827. 9 + 4. 3 + 6 [ ] A [ ] 167129387 (2004) 0320037205,,,, [1, 2 ],, [1, 2 ], 1 ;,,,, ;, ( ), 1 g,,,, CO 2,,, 37, 5% CO 2,, 37,,,,,, 2 1,, Ξ [ ] 2003203217 [ ] 863 (2001AA 213081) [ ] (1972- ),,,, [ ] (1956- ),,,,, E2 m ail: ZhangYL @public. xa. sn. cn
38 ( ) 32,,,, ( 1) 1 6 1. 2,,, ; 300; 2. 3,,, ; 350; 3. 6, (γ ) ; 400; 4. 7, ; 150; 5. 8, ; 400; 6. 9,, 2 4, ; 500 P late 1-6 T he appearance of goat m amm ary ep ithelial cells w h ich culture in v itro 1. 2nd passage, the m amm ary ep ithelial cells fo rm ed island mono layer aggregates as clo sed clusters w ho se boundary is clear and the cells com bined clo sely; 300; 2. 3rd passage, the cell cluster type is betw een open cluster and clo sed cluster w ho se boundary is clear but the cells com bined no t very clo sely; 350; 3. 6th passage, som e elongated cells (γ ) ; 400; 4. 7th passage, appearance of the contact rep ressing cells are no t alike; 150; 5. 8th passage, beeh ive2like m amm ary ep ithelial cells; 400; 6. 9th passage, the cell nucleus are circular o r oval w h ich has 2-4 obvious co res; 500
3 : 39 7 12 7. 10, (γ ) ; 500; 8. 12,, ; 500; 9. 9, (γ ) ; 600; 10. 14,,, ; 150; 11. 13, (γ ) ; 600; 12. 10, ; 40 P late 7-12 T he appearance of goat m amm ary ep ithelial cells w h ich culture in v itro 7. 10th passage, the surface of som e cells em erged flo ss2like rise (γ ) ; 500; 8. 2nd passage, Cells look like big round flat cakes; 500; 9. 9th passage, m amm ary ep ithelial cells contain m ilk drop s (γ ) ; 600; 10. 14th passage,m amm ary ep ithelial cells fo rm ed dom e2 like construction w h ich look s like nipp le and it is called m ilk o rb; 150; 11. 13th passage, m amm ary ep ithelial cells are secreting m ilk (γ ) ; 600; 12. 10th passage, m ilk secreted outside the cells flew and becam e net2like construction; 40, (,, 1), ( 2),,
40 ( ) 32 2 3, ( 1 6, 7 12),, ( 1),,, ( 10) ( 9) ; ( 11), ( 12),,,, ( 5) ;, 2 4, ( 6), ( 8) ; ( 3) ( 7) ( 4) 3,,,, 2 :, ;,,,,,,,,,,,,,, 1 [2 ], Pan tschenko [3 ], [1 ], :,, [4 ] :, [5, 6 ] g [6 ] [7 ] 20 70 Bueh ring [8 ],,,,,, ;, 2 4,, ; ; Pan tschenko [3 ],, [9 ],, COMM A 2D [10 ],,, [ ] [ 1 ]. cam P [D ]. :, 1999. [ 2 ]. [D ]. :, 2002. [ 3 ] Pantschenko A G, Yang T J. M itogenic responsiveness of cap rine m amm ary ep ithelial cells to endocrine and cytok ine facto rs [ J ]. Endocrine, 1999, 10 (2) : 123-130. [ 4 ] T erram ani T T, E ton D, Bui P A, et al. H um an m acrovascular endo thelial cells: op tim ization of culture conditions[j ]. In V itro Cell D ev B io l A nim, 2000, 36 (2) : 125-132. [ 5 ] B lum J L,W icha M S. Ro le of the cyto skeleton in lam inin induced m amm ary gene exp ression[j ]. Cell Physio l, 1988, 135 (1) : 13-22. [ 6 ] Streuli C H, B issell M J. Exp ression of extracellular m atrix components is regulated by substratum [J ]. Cell B io l, 1990, 110 (4) : 1405-
3 1415. : 41 [ 7 ] Parry G, Cullen R, Kaetzel C S, et al. Regulation of differentiation and po larized secretion in m amm ary ep ithelial cells m aintained in culture: extracellular m atrix and m em brane po larity influences[j ]. Cell B io l, 1987, 105 (5) : 2043-2051. [ 8 ] Bueh ring G C. Culture of hum an m amm ary ep ithelial cells: keep ing abreast w ith a new m ethod[j ]. J N atl Cancer Inst, 1972, 49 (5) : 1433-1434. [ 9 ] P ickett P B, P itelka D R, H am amo to S T, et al. O ccluding junctions and cell behavio r in p rim ary cultures of no rm al and neop lastic m amm ary gland cells[j ]. Cell B io l, 1975, 66 (2) : 316-332. [ 10 ] D anielson K G, O bo rn C J, D urban E M. Ep ithelial mouse m amm ary cell line exh ibiting no rm al mo rphogenesis in v ivo and functional differentiation in v itro[j ]. P roc N atl A cad Sci U SA, 1984, 81 (12) : 3756-3760. Study on appearance of goat m amm ary ep ithelial cells w h ich cu ltu red in v itro ZHENG Y ue-mao, PENG X in-rong, XU Y ong-p ing, SH IY u-q iang, ZHANG Y ong (Institute of B io2eng ineering, N orthw est S ci2t ech U niversity of A g riculture and Forestry, Yang ling, S haanx i 712100, China) Abstract: Goat m amm ary ep ithelial cell lines w ere ob tained th rough valid cell cu ltu re m ethods and the appearance of these cells w ere studied by ligh t m icro scope. T he resu lts are as fo llow s: T he cells can fo rm clo sed clu sters and open clu sters, the boundary of clo sed clu sters is clear and the cells com b ined clo se,w h ile the boundary of open clu sters is no t clear and the cells com b ined lax, the cells located in a restricted area. F ib rob last and ep ithelial cells grew together in differen t areas and the tw o cellsπboundary is obviou s; m any cavity2like con struction fo rm ed among the cells. T he pure m amm ary ep ithelial cells w ere cu ltu red in v itro th rough single2cell afloat, part of the cells fo rm ed island mono layer aggregates and part of the cells ex sited in free single cell fo rm. M amm ary ep ithelial cells p ropagated and fo rm ed dom e2like con struction w h ich look s like n ipp le and it is called m ilk o rb; m amm ary ep ithelial cells can p roduce and secrete m ilk, the m ilk secreted ou tside the cells flew and becam e net2like con struction. T he goat m amm ary ep ithelial cells con tain differen t cell type,m ajo rity of the cells are sho rt shu ttle2like o r gubo idal, the cells depended on each o ther clo sely, connected w ith each o ther p rop erly, linked and becam e slice w h ich look s like beeh ive; the cell nucleu s are circu lar o r oval w h ich has 224 co res; part of the cells look like b ig round flat cakes; part of the cells are long; the su rface of som e cells em erged flo ss2like rise; appearance of the con tact rep ressing cells are no t alike. Key words: in v itro cu ltu re; goat; m amm ary ep ithelial cells; the appearance of the cells