JPH07178A - Carrier for cell culture - Google Patents
Carrier for cell cultureInfo
- Publication number
- JPH07178A JPH07178A JP5166175A JP16617593A JPH07178A JP H07178 A JPH07178 A JP H07178A JP 5166175 A JP5166175 A JP 5166175A JP 16617593 A JP16617593 A JP 16617593A JP H07178 A JPH07178 A JP H07178A
- Authority
- JP
- Japan
- Prior art keywords
- glycolipid
- carrier
- cell
- cells
- cell culture
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 238000004113 cell culture Methods 0.000 title claims abstract description 12
- 229930186217 Glycolipid Natural products 0.000 claims abstract description 20
- HVCOBJNICQPDBP-UHFFFAOYSA-N 3-[3-[3,5-dihydroxy-6-methyl-4-(3,4,5-trihydroxy-6-methyloxan-2-yl)oxyoxan-2-yl]oxydecanoyloxy]decanoic acid;hydrate Chemical compound O.OC1C(OC(CC(=O)OC(CCCCCCC)CC(O)=O)CCCCCCC)OC(C)C(O)C1OC1C(O)C(O)C(O)C(C)O1 HVCOBJNICQPDBP-UHFFFAOYSA-N 0.000 claims abstract description 17
- 125000002252 acyl group Chemical group 0.000 claims abstract description 5
- 125000004432 carbon atom Chemical group C* 0.000 claims abstract description 5
- 239000000126 substance Substances 0.000 claims abstract description 5
- 230000021164 cell adhesion Effects 0.000 claims description 4
- 230000001737 promoting effect Effects 0.000 claims description 3
- 210000004027 cell Anatomy 0.000 description 27
- 238000001000 micrograph Methods 0.000 description 15
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 12
- 238000000034 method Methods 0.000 description 8
- 230000000052 comparative effect Effects 0.000 description 7
- 235000014113 dietary fatty acids Nutrition 0.000 description 7
- 229930195729 fatty acid Natural products 0.000 description 7
- 239000000194 fatty acid Substances 0.000 description 7
- 150000004665 fatty acids Chemical class 0.000 description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- 102000008186 Collagen Human genes 0.000 description 5
- 108010035532 Collagen Proteins 0.000 description 5
- 229920001436 collagen Polymers 0.000 description 5
- 235000011187 glycerol Nutrition 0.000 description 5
- 230000001070 adhesive effect Effects 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 230000035755 proliferation Effects 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 239000000853 adhesive Substances 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 150000002339 glycosphingolipids Chemical class 0.000 description 3
- WRIDQFICGBMAFQ-UHFFFAOYSA-N (E)-8-Octadecenoic acid Natural products CCCCCCCCCC=CCCCCCCC(O)=O WRIDQFICGBMAFQ-UHFFFAOYSA-N 0.000 description 2
- LQJBNNIYVWPHFW-UHFFFAOYSA-N 20:1omega9c fatty acid Natural products CCCCCCCCCCC=CCCCCCCCC(O)=O LQJBNNIYVWPHFW-UHFFFAOYSA-N 0.000 description 2
- QSBYPNXLFMSGKH-UHFFFAOYSA-N 9-Heptadecensaeure Natural products CCCCCCCC=CCCCCCCCC(O)=O QSBYPNXLFMSGKH-UHFFFAOYSA-N 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- ZQPPMHVWECSIRJ-UHFFFAOYSA-N Oleic acid Natural products CCCCCCCCC=CCCCCCCCC(O)=O ZQPPMHVWECSIRJ-UHFFFAOYSA-N 0.000 description 2
- 239000005642 Oleic acid Substances 0.000 description 2
- 238000007605 air drying Methods 0.000 description 2
- 210000004102 animal cell Anatomy 0.000 description 2
- 239000012091 fetal bovine serum Substances 0.000 description 2
- IPCSVZSSVZVIGE-UHFFFAOYSA-N hexadecanoic acid Chemical compound CCCCCCCCCCCCCCCC(O)=O IPCSVZSSVZVIGE-UHFFFAOYSA-N 0.000 description 2
- QXJSBBXBKPUZAA-UHFFFAOYSA-N isooleic acid Natural products CCCCCCCC=CCCCCCCCCC(O)=O QXJSBBXBKPUZAA-UHFFFAOYSA-N 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- ZQPPMHVWECSIRJ-KTKRTIGZSA-N oleic acid Chemical compound CCCCCCCC\C=C/CCCCCCCC(O)=O ZQPPMHVWECSIRJ-KTKRTIGZSA-N 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 239000012679 serum free medium Substances 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 235000021122 unsaturated fatty acids Nutrition 0.000 description 2
- 150000004670 unsaturated fatty acids Chemical class 0.000 description 2
- OYHQOLUKZRVURQ-NTGFUMLPSA-N (9Z,12Z)-9,10,12,13-tetratritiooctadeca-9,12-dienoic acid Chemical compound C(CCCCCCC\C(=C(/C\C(=C(/CCCCC)\[3H])\[3H])\[3H])\[3H])(=O)O OYHQOLUKZRVURQ-NTGFUMLPSA-N 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 241000699800 Cricetinae Species 0.000 description 1
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 1
- 102000016359 Fibronectins Human genes 0.000 description 1
- 108010067306 Fibronectins Proteins 0.000 description 1
- PNNNRSAQSRJVSB-SLPGGIOYSA-N Fucose Natural products C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)C=O PNNNRSAQSRJVSB-SLPGGIOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- SHZGCJCMOBCMKK-DHVFOXMCSA-N L-fucopyranose Chemical compound C[C@@H]1OC(O)[C@@H](O)[C@H](O)[C@@H]1O SHZGCJCMOBCMKK-DHVFOXMCSA-N 0.000 description 1
- 102000007547 Laminin Human genes 0.000 description 1
- 108010085895 Laminin Proteins 0.000 description 1
- 102000004882 Lipase Human genes 0.000 description 1
- 108090001060 Lipase Proteins 0.000 description 1
- 239000004367 Lipase Substances 0.000 description 1
- OVRNDRQMDRJTHS-CBQIKETKSA-N N-Acetyl-D-Galactosamine Chemical compound CC(=O)N[C@H]1[C@@H](O)O[C@H](CO)[C@H](O)[C@@H]1O OVRNDRQMDRJTHS-CBQIKETKSA-N 0.000 description 1
- OVRNDRQMDRJTHS-UHFFFAOYSA-N N-acelyl-D-glucosamine Natural products CC(=O)NC1C(O)OC(CO)C(O)C1O OVRNDRQMDRJTHS-UHFFFAOYSA-N 0.000 description 1
- MBLBDJOUHNCFQT-UHFFFAOYSA-N N-acetyl-D-galactosamine Natural products CC(=O)NC(C=O)C(O)C(O)C(O)CO MBLBDJOUHNCFQT-UHFFFAOYSA-N 0.000 description 1
- OVRNDRQMDRJTHS-FMDGEEDCSA-N N-acetyl-beta-D-glucosamine Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O OVRNDRQMDRJTHS-FMDGEEDCSA-N 0.000 description 1
- MBLBDJOUHNCFQT-LXGUWJNJSA-N N-acetylglucosamine Natural products CC(=O)N[C@@H](C=O)[C@@H](O)[C@H](O)[C@H](O)CO MBLBDJOUHNCFQT-LXGUWJNJSA-N 0.000 description 1
- 235000021314 Palmitic acid Nutrition 0.000 description 1
- 239000004698 Polyethylene Substances 0.000 description 1
- 108010039918 Polylysine Proteins 0.000 description 1
- 239000004793 Polystyrene Substances 0.000 description 1
- 235000021355 Stearic acid Nutrition 0.000 description 1
- 239000003377 acid catalyst Substances 0.000 description 1
- 230000010933 acylation Effects 0.000 description 1
- 238000005917 acylation reaction Methods 0.000 description 1
- 230000001464 adherent effect Effects 0.000 description 1
- 125000003158 alcohol group Chemical group 0.000 description 1
- WQZGKKKJIJFFOK-PHYPRBDBSA-N alpha-D-galactose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-PHYPRBDBSA-N 0.000 description 1
- DTOSIQBPPRVQHS-PDBXOOCHSA-N alpha-linolenic acid Chemical compound CC\C=C/C\C=C/C\C=C/CCCCCCCC(O)=O DTOSIQBPPRVQHS-PDBXOOCHSA-N 0.000 description 1
- 235000020661 alpha-linolenic acid Nutrition 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 230000004956 cell adhesive effect Effects 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 229930182830 galactose Natural products 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 125000001165 hydrophobic group Chemical group 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 229960004488 linolenic acid Drugs 0.000 description 1
- KQQKGWQCNNTQJW-UHFFFAOYSA-N linolenic acid Natural products CC=CCCC=CCC=CCCCCCCCC(O)=O KQQKGWQCNNTQJW-UHFFFAOYSA-N 0.000 description 1
- 235000019421 lipase Nutrition 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- WQEPLUUGTLDZJY-UHFFFAOYSA-N n-Pentadecanoic acid Natural products CCCCCCCCCCCCCCC(O)=O WQEPLUUGTLDZJY-UHFFFAOYSA-N 0.000 description 1
- 229950006780 n-acetylglucosamine Drugs 0.000 description 1
- 231100000587 neutral red assay Toxicity 0.000 description 1
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 description 1
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 description 1
- 235000021313 oleic acid Nutrition 0.000 description 1
- 150000002482 oligosaccharides Polymers 0.000 description 1
- -1 polyethylene Polymers 0.000 description 1
- 229920000573 polyethylene Polymers 0.000 description 1
- 229920000656 polylysine Polymers 0.000 description 1
- 229920002223 polystyrene Polymers 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 150000004671 saturated fatty acids Chemical class 0.000 description 1
- 239000008117 stearic acid Substances 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
Landscapes
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Description
【０００１】[0001]
【産業上の利用分野】本発明は、細胞培養用担体に関す
る。TECHNICAL FIELD The present invention relates to a carrier for cell culture.
【０００２】[0002]
【従来の技術】従来、動物細胞の培養には牛胎仔血清添
加培地が使用されてきたが、当該培地は高価であり、且
つロット間のばらつきがあることから、血清を使用しな
い無血清培地の利用が増加しつつある。2. Description of the Related Art Conventionally, a medium containing fetal bovine serum has been used for culturing animal cells. However, since the medium is expensive and varies among lots, a serum-free medium containing no serum is used. Usage is increasing.
【０００３】しかし、接着性細胞を無血清培養するに
は、フィブロネクチン、ラミニン、コラーゲンやポリリ
ジンといった接着因子を必要とし、これらを添加した培
地や、予めこれらの接着因子で表面をコーティングした
組織培養器具の利用が必要となっている。However, in order to culture adherent cells without serum, adhesion factors such as fibronectin, laminin, collagen and polylysine are required, and a culture medium to which these are added or a tissue culture instrument whose surface is previously coated with these adhesion factors. Is required.
【０００４】又、オリゴ糖鎖やスフィンゴ糖脂質は、細
胞間の特異的相互作用発現に関与していると考えられて
おり、これらの物質が細胞接着効果を有していることも
実験的に証明されている。Further, oligosaccharide chains and glycosphingolipids are considered to be involved in the expression of specific interactions between cells, and it has been experimentally shown that these substances have a cell adhesion effect. Proven.
【０００５】更に、培養器具の素材を改良し、器具の表
面に電荷を与えることにより、接着効果をもたせる方法
も実用化されている。Further, a method of improving the material of the culture instrument and giving an electric charge to the surface of the instrument to give an adhesive effect has been put into practical use.
【０００６】しかしながら、これらのタンパク質やスフ
ィンゴ糖脂質等の接着因子はいずれも高価格であった
り、培養器具の改良単独では効果が不十分であった。However, the adhesion factors such as these proteins and glycosphingolipids are all expensive, and the improvement of the culture device alone is not effective enough.
【０００７】[0007]
【発明が解決しようとする課題】本発明は、コラーゲン
等のタンパク性接着因子、スフィンゴ糖脂質系の接着因
子と比べて非常に安価であり、かつ調製が容易な細胞培
養用担体を提供することを目的になされたものである。DISCLOSURE OF THE INVENTION The present invention provides a carrier for cell culture, which is much cheaper and easier to prepare than proteinaceous adhesion factors such as collagen and glycosphingolipid-based adhesion factors. It was made for the purpose.
【０００８】[0008]
【課題を解決するための手段】本発明者らは、上記課題
を達成すべく鋭意検討の結果、特定の構造を有する糖脂
質にコラーゲン等の接着性タンパク質に相当する細胞接
着作用のあることを見いだし、かかる知見に基づいて本
発明を完成するに至った。Means for Solving the Problems As a result of intensive studies to achieve the above object, the present inventors have found that glycolipids having a specific structure have a cell adhesive action equivalent to an adhesive protein such as collagen. The present invention has been completed and the present invention has been completed based on such findings.
【０００９】即ち、本発明に係る細胞培養用担体は、糖
脂質、特に炭素数８〜２２のアシル基を有するグリセロ
糖脂質を細胞接着促進物質とすることを特徴とする。That is, the carrier for cell culture according to the present invention is characterized by using a glycolipid, particularly a glyceroglycolipid having an acyl group having 8 to 22 carbon atoms as a cell adhesion promoting substance.
【００１０】本発明で用いられるグリセロ糖脂質は、分
子内に糖とグリセリンとがグリコシド結合しており、こ
の分子に脂肪酸がエステル結合した構造を有する。The glyceroglycolipid used in the present invention has a structure in which sugar and glycerin are glycosidic-bonded in the molecule, and fatty acid is ester-bonded to this molecule.
【００１１】かかるグリセロ糖脂質は、例えば酸触媒の
存在下、糖とグリセリンより糖グリセロールを調製し、
ついで適当な脂肪酸を用いてアシル化することにより得
ることができる。更には、先に本出願人が提案したリパ
ーゼを用いる糖脂質の合成方法（特願平３−１０２０８
２号）が好適に採用される。Such glyceroglycolipid is prepared by preparing sugar glycerol from sugar and glycerin in the presence of an acid catalyst,
Then, it can be obtained by acylation with a suitable fatty acid. Furthermore, a method for synthesizing glycolipids using lipase previously proposed by the present applicant (Japanese Patent Application No. 3-10208).
No. 2) is preferably adopted.
【００１２】当該糖としては、例えばグルコース、ガラ
クトース、マンノース、フコース、Ｎ−アセチルグルコ
サミン、Ｎ−アセチルガラクトサミン等の単糖が挙げら
れる。一般に動物細胞には種特異性、臓器特異性が存在
するため、培養する細胞に親和性の強い糖鎖を選択する
ことが可能である。Examples of the sugar include monosaccharides such as glucose, galactose, mannose, fucose, N-acetylglucosamine and N-acetylgalactosamine. Since animal cells generally have species-specificity and organ-specificity, it is possible to select a sugar chain having a strong affinity for cells to be cultured.
【００１３】脂肪酸としては、炭素数８〜２２の飽和若
しくは不飽和の脂肪酸であり、特に炭素数１２〜２０程
度の脂肪酸が好ましい。当該脂肪酸として、具体的に
は、パルミチン酸、ステアリン酸、オレイン酸、リノー
ル酸、リノレン酸等が例示される。中でも上記炭素数を
有する不飽和脂肪酸が推奨される。The fatty acid is a saturated or unsaturated fatty acid having 8 to 22 carbon atoms, and a fatty acid having about 12 to 20 carbon atoms is particularly preferable. Specific examples of the fatty acid include palmitic acid, stearic acid, oleic acid, linoleic acid, and linolenic acid. Among them, unsaturated fatty acids having the above carbon number are recommended.
【００１４】本発明に係る糖脂質としては、１個のアシ
ル基を有する糖脂質が推奨されるが、このものに２個以
上のアシル基を有する糖脂質が混在していてもよい。As the glycolipid according to the present invention, a glycolipid having one acyl group is recommended, but a glycolipid having two or more acyl groups may be mixed therein.
【００１５】脂肪酸がエステル結合している部位は、糖
グリセロール中のいずれでもよいが、好ましくはグリセ
リンの末端アルコール若しくは糖の６位がよい。又、エ
ステル結合の部位が一定していない混合物を使用するこ
とも可能である。The site where the fatty acid is ester-bonded may be any position in the sugar glycerol, but is preferably the terminal alcohol of glycerin or the 6-position of the sugar. It is also possible to use a mixture in which the sites of ester bonds are not constant.
【００１６】このようにして得られた糖脂質を担体に利
用するには、種々の方法がある。例えば、糖脂質をエタ
ノール等の溶剤に溶かし、この溶液を細胞培養容器に加
え、溶剤を風乾して除くことにより、糖脂質でコーティ
ングされた細胞培養容器を得ることができる。There are various methods for utilizing the glycolipid thus obtained as a carrier. For example, a glycolipid-coated cell culture vessel can be obtained by dissolving a glycolipid in a solvent such as ethanol, adding this solution to the cell culture vessel, and air-drying the solvent.
【００１７】使用する糖脂質の濃度としては、１〜１０
０μg/cm2程度が良く、より望ましくは５〜５０μg/cm2
程度である。The concentration of glycolipid used is 1 to 10
0 Pg / cm 2 about well, more preferably 5-50 [mu] g / cm 2
It is a degree.
【００１８】使用する培養器具の素材はいずれでも良い
が、糖脂質の疎水基が吸着しやすいようなポリスチレ
ン、ポリエチレン等の樹脂が好ましい。Any material may be used for the culture instrument to be used, but a resin such as polystyrene, polyethylene or the like is preferred because the hydrophobic group of the glycolipid is easily adsorbed.
【００１９】[0019]
【実施例】以下に実施例を挙げて本発明を詳しく説明す
る。尚、担体へのコーティングは、各濃度の糖脂質のエ
タノール溶液１００μlをマルチウェルプレート上に注
ぎ静置、風乾することにより行った。EXAMPLES The present invention will be described in detail below with reference to examples. The carrier was coated by pouring 100 μl of an ethanol solution of glycolipid at each concentration onto a multiwell plate, allowing it to stand, and air-drying.
【００２０】実施例１ ハムスター肺ＣＨＬ／ＩＵ細胞（およそ２×１０4）を
懸濁させた成長因子添加無血清培地０．５mlを、脂肪酸
としてオレイン酸を選択して調製されるグリセロ糖脂質
（濃度１μg/cm2）でプレコートした細胞培養用マルチ
ウェルプレート(48well)に加え、５％ＣＯ2存在下、３
７℃で７日間培養した。この細胞の成長状態の顕微鏡観
察下での写真を図１に示す。更に、この細胞数をニュー
トラルレッドアッセイ法により測定したところ、３×１
０3であった。Example 1 Glyceroglycolipids prepared by selecting oleic acid as a fatty acid in 0.5 ml of a serum-free medium containing growth factors in which hamster lung CHL / IU cells (about 2 × 10 4 ) were suspended ( Add to a multi-well plate for cell culture (48 well) pre-coated with a concentration of 1 μg / cm 2 ) and in the presence of 5% CO 2 ,
It was cultured at 7 ° C for 7 days. A photograph of the growth state of the cells under a microscope is shown in FIG. Furthermore, when the number of cells was measured by the neutral red assay method, it was 3 × 1.
It was 0 3 .
【００２１】実施例２ グリセロ糖脂質濃度を５μg/cm2とした他は実施例１と
同様にして培養した。このとき得られた細胞状態の顕微
鏡写真を図２に示す。又、この細胞数を実施例１と同様
にして測定したところ、３×１０4であった。Example 2 Culture was performed in the same manner as in Example 1 except that the glyceroglycolipid concentration was 5 μg / cm 2 . A micrograph of the cell state obtained at this time is shown in FIG. When the number of cells was measured in the same manner as in Example 1, it was 3 × 10 4 .
【００２２】実施例３ グリセロ糖脂質濃度を１０μg/cm2とした他は実施例１
と同様にして培養した。このとき得られた細胞状態の顕
微鏡写真を図３に示す。又、この細胞数を実施例１と同
様にして測定したところ、６×１０4であった。Example 3 Example 1 except that the glyceroglycolipid concentration was 10 μg / cm 2.
It culture | cultivated similarly to. A micrograph of the cell state obtained at this time is shown in FIG. When the number of cells was measured in the same manner as in Example 1, it was 6 × 10 4 .
【００２３】比較例１ 糖脂質でプレコートしないプレートを用いて培養した他
は実施例１と同様にして培養した。このとき得られた細
胞状態の顕微鏡写真を図４に示す。又、この細胞数を実
施例１と同様にして測定したところ、１×１０3であっ
た。Comparative Example 1 Culture was carried out in the same manner as in Example 1 except that the plate was not precoated with glycolipid. A micrograph of the cell state obtained at this time is shown in FIG. When the number of cells was measured in the same manner as in Example 1, it was 1 × 10 3 .
【００２４】比較例２ １０％牛胎仔血清含有培地（Eagle's minimal essentia
l medium）を用いて培養した他は実施例１と同様にして
培養した。このとき得られた細胞状態の顕微鏡写真を図
５に示す。又、この細胞数を実施例１と同様にして測定
したところ、２×１０5であった。Comparative Example 2 Medium containing 10% fetal bovine serum (Eagle's minimal essentia
The culture was performed in the same manner as in Example 1 except that the medium was used. A micrograph of the cell state obtained at this time is shown in FIG. When the number of cells was measured in the same manner as in Example 1, it was 2 × 10 5 .
【００２５】比較例３ ０．０３％コラーゲンでプレコートしたプレートを用い
た他は実施例１と同様にして培養した。このとき得られ
た細胞状態の顕微鏡写真を図６に示す。又、この細胞数
を実施例１と同様にして測定したところ、６×１０3で
あった。Comparative Example 3 Culture was performed in the same manner as in Example 1 except that a plate precoated with 0.03% collagen was used. A micrograph of the cell state obtained at this time is shown in FIG. The number of cells was measured in the same manner as in Example 1 and found to be 6 × 10 3 .
【００２６】[0026]
【発明の効果】本発明に係る糖脂質は、従来用いられて
きた接着性タンパク質と遜色のない細胞の接着効果を示
す。この糖脂質をコーティングして得られる細胞培養用
担体はより安価に調製が可能であり、コラーゲン等の接
着性タンパク質に代わる細胞の接着促進物質としての担
体として有効に利用することができる。EFFECTS OF THE INVENTION The glycolipid according to the present invention exhibits a cell-adhesion effect comparable to that of the conventionally used adhesive protein. The carrier for cell culture obtained by coating with this glycolipid can be prepared at a lower cost, and can be effectively used as a carrier as an adhesion promoting substance for cells in place of an adhesive protein such as collagen.
【００２７】[0027]
【図１】[Figure 1]
【図２】[Fig. 2]
【図３】[Figure 3]
【図４】[Figure 4]
【図５】[Figure 5]
【００２８】[0028]
第１図は、実施例１で得られたＣＨＬ／ＩＵ細胞の増殖
状態を示す顕微鏡写真（１００倍）である。FIG. 1 is a micrograph (100 ×) showing the growth state of CHL / IU cells obtained in Example 1.
【００２９】第２図は、実施例２で得られたＣＨＬ／Ｉ
Ｕ細胞の増殖状態を示す顕微鏡写真（１００倍）であ
る。FIG. 2 shows the CHL / I obtained in Example 2.
It is a micrograph (100 times) which shows the proliferation state of U cell.
【００３０】第３図は、実施例３で得られたＣＨＬ／Ｉ
Ｕ細胞の増殖状態を示す顕微鏡写真（１００倍）であ
る。FIG. 3 shows the CHL / I obtained in Example 3.
It is a micrograph (100 times) which shows the proliferation state of U cell.
【００３１】第４図は、比較例１で得られたＣＨＬ／Ｉ
Ｕ細胞の増殖状態を示す顕微鏡写真（１００倍）であ
る。FIG. 4 shows the CHL / I obtained in Comparative Example 1.
It is a micrograph (100 times) which shows the proliferation state of U cell.
【００３２】第５図は、比較例２で得られたＣＨＬ／Ｉ
Ｕ細胞の増殖状態を示す顕微鏡写真（１００倍）であ
る。FIG. 5 shows the CHL / I obtained in Comparative Example 2.
It is a micrograph (100 times) which shows the proliferation state of U cell.
【手続補正書】[Procedure amendment]
【提出日】平成６年５月２７日[Submission date] May 27, 1994
【手続補正１】[Procedure Amendment 1]
【補正対象書類名】明細書[Document name to be amended] Statement
【補正対象項目名】００２７[Name of item to be corrected] 0027
【補正方法】削除[Correction method] Delete
【手続補正２】[Procedure Amendment 2]
【補正対象書類名】明細書[Document name to be amended] Statement
【補正対象項目名】図面の簡単な説明[Name of item to be corrected] Brief description of the drawing
【補正方法】変更[Correction method] Change
【補正内容】[Correction content]
【図面の簡単な説明】[Brief description of drawings]
【図１】 図１は実施例１で得られたＣＨＬ／ＩＵ細胞
の増殖状態を示す顕微鏡写真（１００倍）である。FIG. 1 is a micrograph (100 ×) showing the growth state of CHL / IU cells obtained in Example 1.
【図２】 図２は実施例２で得られたＣＨＬ／ＩＵ細胞
の増殖状態を示す顕微鏡写真（１００倍）である。FIG. 2 is a micrograph (100 ×) showing the growth state of CHL / IU cells obtained in Example 2.
【図３】 図３は実施例３で得られたＣＨＬ／ＩＵ細胞
の増殖状態を示す顕微鏡写真（１００倍）である。FIG. 3 is a micrograph (100 ×) showing the growth state of CHL / IU cells obtained in Example 3.
【図４】 図４は比較例１で得られたＣＨＬ／ＩＵ細胞
の増殖状態を示す顕微鏡写真（１００倍）である。FIG. 4 is a micrograph (100 ×) showing the growth state of CHL / IU cells obtained in Comparative Example 1.
【図５】 図５は比較例２で得られたＣＨＬ／ＩＵ細胞
の増殖状態を示す顕微鏡写真（１００倍）である。FIG. 5 is a micrograph (100 ×) showing the growth state of CHL / IU cells obtained in Comparative Example 2.
Claims (2)
特徴とする細胞培養用担体。1. A carrier for cell culture, which comprises a glycolipid as a cell adhesion promoting substance.
するグリセロ糖脂質である請求項１に記載の細胞培養用
担体。2. The carrier for cell culture according to claim 1, wherein the glycolipid is a glyceroglycolipid having an acyl group having 8 to 22 carbon atoms.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP5166175A JPH07178A (en) | 1993-06-11 | 1993-06-11 | Carrier for cell culture |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP5166175A JPH07178A (en) | 1993-06-11 | 1993-06-11 | Carrier for cell culture |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH07178A true JPH07178A (en) | 1995-01-06 |
Family
ID=15826473
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP5166175A Pending JPH07178A (en) | 1993-06-11 | 1993-06-11 | Carrier for cell culture |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH07178A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2003074099A1 (en) * | 2002-03-06 | 2003-09-12 | Japan Tissue Engineering Co.,Ltd | Base material for tissue regeneration, transplantation material and process for producing the same |
| WO2003074691A1 (en) * | 2002-03-01 | 2003-09-12 | National Institute Of Advanced Industrial Science And Technology | Immobilized cells and liposomes and method of immobilizing the same |
| JP2010187680A (en) * | 1998-11-19 | 2010-09-02 | Organogenesis Inc | Bioengineered tissue construct and method for producing and using the same |
| JP2010536350A (en) * | 2007-08-20 | 2010-12-02 | エバーハルト・カールス・ユニバーシタット テュービンゲン ユニバーシタットスクリニクム | Collagen-containing cell carrier |
-
1993
- 1993-06-11 JP JP5166175A patent/JPH07178A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2010187680A (en) * | 1998-11-19 | 2010-09-02 | Organogenesis Inc | Bioengineered tissue construct and method for producing and using the same |
| WO2003074691A1 (en) * | 2002-03-01 | 2003-09-12 | National Institute Of Advanced Industrial Science And Technology | Immobilized cells and liposomes and method of immobilizing the same |
| US7501280B2 (en) | 2002-03-01 | 2009-03-10 | National Institute Of Advanced Industrial Science And Technology | Immobilized cells and liposomes and method of immobilizing the same |
| WO2003074099A1 (en) * | 2002-03-06 | 2003-09-12 | Japan Tissue Engineering Co.,Ltd | Base material for tissue regeneration, transplantation material and process for producing the same |
| JP2010536350A (en) * | 2007-08-20 | 2010-12-02 | エバーハルト・カールス・ユニバーシタット テュービンゲン ユニバーシタットスクリニクム | Collagen-containing cell carrier |
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