JPWO2020170943A1 - 脱細胞化処理液及び洗浄組成物 - Google Patents
脱細胞化処理液及び洗浄組成物 Download PDFInfo
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- JPWO2020170943A1 JPWO2020170943A1 JP2020571729A JP2020571729A JPWO2020170943A1 JP WO2020170943 A1 JPWO2020170943 A1 JP WO2020170943A1 JP 2020571729 A JP2020571729 A JP 2020571729A JP 2020571729 A JP2020571729 A JP 2020571729A JP WO2020170943 A1 JPWO2020170943 A1 JP WO2020170943A1
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Abstract
Description
界面活性剤はタンパク質、脂質などの物質を非選択的に除去するため、界面活性剤を用いて脱細胞化処理を行った生体組織では、細胞外マトリックス成分を構成するたんぱく質が劣化しており、脱細胞化組織の強度劣化や細胞の再接着性能悪化につながり、生体適合性が低い。そのため界面活性剤の一種であるソホロースリピッドを使用して脱細胞化処理を行った場合も、他の界面活性剤と同様に、細胞外マトリックス成分を構成するたんぱく質が劣化し、生体適合性が低いものと予想される。しかしながら本発明者はこの予想に反してソホロースリピッドを動物由来組織の脱細胞化に用いた場合、細胞のみを除去し、細胞外マトリックス成分を構成するたんぱく質を劣化させることなく組織の脱細胞化が可能であることを新知見として見出し、かかる事実に基づいて本発明を完成させた。
脱細胞化処理を行った生体組織には、組織の残分やその脱細胞化処理に使用した界面活性剤などの物質が残存しているため、脱細胞化組織を再細胞化する前に残存している物質を洗浄することが好ましい。
下記に本実施形態にかかる脱細胞化処理液を使用する脱細胞化組織の製造方法の一実施形態を説明する。
超純水、10重量%、20重量%のSL溶液中にラット新生仔の肺を浸漬し、振とうした。1日後、2日後、5日後、10日後に実体顕微鏡を用いて脱細胞化効果を確認した。結果を表1及び図1に示す。
0.1 重量%、1.0 重量%、10 重量%のSL溶液、SDS(ラウリル硫酸ナトリウム) 溶液、Triton 溶液、APG(アルキルポリグリコシド) 溶液中にラット新生仔の肺を浸漬し、振とうした。2日、4日、6日後に脱細胞化状態として実体顕微鏡を用いて透明性の割合で評価、細胞外マトリックスへの影響として目視により臓器の縮小状態(大きさ)や臓器の崩壊状態、ピンセットで掴んだ際の強度で総合的に評価した。また、より詳細な細胞外マトリックスへの影響評価として、共焦点レーザー顕微鏡を用いての直接観察及び細胞外マトリックスの主要成分であるコラーゲンの免疫染色により内部構造への影響を確認した。脱細胞化状態の評価結果を表2−1,表2−2,表2−3に示す。細胞外マトリックスへの影響の評価結果を表3−1,表3−2,表3−3に示す。表3の結果は表2の非常に良好になった日数で確認した。図2の写真は浸漬後6日目の写真である。
各処理液で脱細胞化処理後の血管について、1 次抗体:Anti-CollagenIV、2 次抗体: Anti-Rabbit 488 を用いて免疫染色を行い観察した。また血管の内腔について面積測定により評価し、細胞外マトリックスへの影響を確認した。免疫染色の観察写真を図9、図10に、面積測定結果を図11、図12に示した。
超純水、10重量%、20重量%のSL溶液中にラット新生仔の肝臓を浸漬し、振とうした。1日後、2日後、5日後、10日後に実体顕微鏡を用いて脱細胞化効果を確認した。結果を表4及び図13に示す。
超純水、1重量%、10重量%、20重量%のSL溶液中にラット新生仔の心臓を浸漬し、振とうした。1日後、2日後、5日後、10日後に実体顕微鏡を用いて脱細胞化効果を確認した。結果を表5及び図14に示す。
超純水、10重量%、20重量%のSL溶液中にラット新生仔の腎臓を浸漬し、振とうした。1日後、2日後、5日後、10日後に実体顕微鏡を用いて脱細胞化効果を確認した。結果を表6及び図15に示す。
超純水、0.1重量%、1重量%、10重量%、20重量%のSL溶液中にラット新生仔の皮膚を浸漬し、振とうした。1日後、2日後、5日後、10日後に実体顕微鏡を用いて脱細胞化効果を確認した。結果を表9及び図17に示す。
超純水、0.1重量%、1重量%、10重量%、20重量%のSL溶液中にラット新生仔の腸を浸漬し、振とうした。1日後、2日後、5日後、10日後に実体顕微鏡を用いて脱細胞化効果を確認した。結果を表12及び図19に示す。
超純水、1重量%、10重量%、20重量%のSL溶液中にラット新生仔の脾臓を浸漬し、振とうした。1日後、2日後、5日後、10日後に実体顕微鏡を用いて脱細胞化効果を確認した。結果を表13及び図20に示す。
ラット新生仔心臓を摘出し、室温において溶液に含浸・振盪処理を5日間行った。実体顕微鏡にて透過光と実体画像により、心臓の透明性を確認することで脱細胞化及び形状維持を目視にて評価した。結果を表16及び図22に示す。図22において、同じ心臓を上段は実体画像による観察した写真図であり、下段は透過光による観察した写真図である。
ラット新生仔の心臓を25℃(室温)で溶液中に24時間含浸・振盪処理後、細胞外マトリックス構成蛋白(フィブロネクチン)の存在と分布を蛍光実体顕微鏡、共焦点顕微鏡で観察した。結果を表17及び図23に示す。
ラット新生仔の心臓を25℃(室温)で各溶液に24時間含浸・振盪処理後の心臓を6時間洗浄した。その後、新生仔ラット心筋細胞(2 x 106)を上から添加培養した。播種7日目の心臓の拍動を形態観察により確認した。
ラット新生仔の心臓を25℃(室温)で1%SDSに24時間含浸・振盪処理後の心臓を表11の組成にて6時間洗浄した。その後、新生仔ラット心筋細胞(2 x 106)を上から添加培養した。播種7日目の心臓の拍動を確認した。
ラット新生仔の心臓を25℃(室温)で1%SDS+0.5%SLに24時間含浸・振盪処理後の心臓を表20の組成にて6時間洗浄した。その後、新生仔ラット心筋細胞(2x106)を上から添加培養した。播種7日目の心臓の拍動を確認した。
ラット新生仔の心臓を25℃(室温)で表21に示す溶液中に24時間含浸・振盪処理後、心臓の容積比を観察した。結果を表21及び図24に示す。図24において、同じ心臓を上段は実体画像による観察した写真図であり、下段は透過光による観察した写真図である。
ラット新生仔の心臓を25℃(室温)で1.0%SLに24時間含浸・振盪処理後、蛍光顕微鏡により観察をした。図25上段に示されるように、collagen type1、type4 ともに生体心臓と同様の鱗状構造が保持されていた。また、α-actinin 陽性細胞(心筋細胞)は検出されず、脱細胞化が確認された。
Claims (6)
- 動物由来組織の脱細胞化に用いる脱細胞化処理液であり、0.01重量%以上20重量%以下のソホロースリピッド含む脱細胞化処理液。
- 更に0.1重量%以上10重量%以下の界面活性剤を含む請求項1に記載の脱細胞化処理液。
- 前記界面活性剤はラウリル硫酸ナトリウムである請求項2記載の脱細胞化処理液。
- 0.1重量%以上1.0重量%以下のソホロースリピッドを含むとともに0.1重量%以上1.0重量%以下のラウリル硫酸ナトリウムを包含する請求項3に記載の脱細胞化処理液。
- 脱細胞化処理後の動物由来組織の洗浄に使用する洗浄組成物であり、0.01重量%以上10重量%以下のソホロースリピッドを含む洗浄組成物。
- 酢酸緩衝液、リン酸緩衝液、クエン酸緩衝液、ホウ酸緩衝液、酒石酸緩衝液、トリス緩衝液、HEPES緩衝液及びMES緩衝液から少なくとも一つを含む緩衝液を含む請求項5に記載の洗浄組成物。
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Non-Patent Citations (8)
Title |
---|
CELL, vol. 157, JPN6020017073, 2014, pages 726 - 739, ISSN: 0004471722 * |
NAT. MED., vol. 14, no. 2, JPN6021005198, 2008, pages 213 - 221, ISSN: 0004471727 * |
NAT. MED., vol. 16, no. 7, JPN6021005197, 2010, pages 814 - 820, ISSN: 0004471726 * |
NAT. MED., vol. 16, no. 8, JPN6021005196, 2010, pages 927 - 933, ISSN: 0004471725 * |
PLOS ONE, vol. 12(7), JPN7020001339, 2017, pages 0182072, ISSN: 0004471723 * |
SCIENCE, vol. 329, JPN7021000510, 2010, pages 538 - 541, ISSN: 0004471724 * |
TISSUE ENG. PART A, vol. 16, no. 7, JPN7021000511, 2010, pages 2207 - 2216, ISSN: 0004471728 * |
再生医療, vol. 10, no. 3, JPN7020001338, 2011, pages 132 - 135, ISSN: 0004471721 * |
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