CN117018295A - 一种成活率高的自体脂肪移植方法 - Google Patents
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- CN117018295A CN117018295A CN202311081095.XA CN202311081095A CN117018295A CN 117018295 A CN117018295 A CN 117018295A CN 202311081095 A CN202311081095 A CN 202311081095A CN 117018295 A CN117018295 A CN 117018295A
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
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- A61L27/3895—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells using specific culture conditions, e.g. stimulating differentiation of stem cells, pulsatile flow conditions
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Abstract
本发明涉及脂肪移植技术领域,尤其涉及一种成活率高的自体脂肪移植方法。所述方案包括以下步骤:1)提取脂肪干细胞;2)用血管内皮细胞培养得到的上清液处理脂肪干细胞;3)将步骤2)中经处理的脂肪干细胞与颗粒脂肪混合,进行脂肪移植。所述上清液是血管内皮细胞在低血清、低氧和低一氧化氮的条件下培养得到的。本发明制备的胁迫状态下内皮细胞上清液对于OGD条件下的脂肪干细胞的活性具有良好的保持作用;可以提高移植物最终的体积和质量保留率;可以抑制脂肪移植物凋亡;显著提高移植物新血管生成的能力;促进脂肪移植物的成活率。
Description
技术领域
本发明涉及脂肪移植技术领域,尤其涉及一种成活率高的自体脂肪移植方法。
背景技术
组织移植是整形外科常用的外科手段之一,用以修复畸形、外伤、肿瘤等造成的畸形和组织缺损,其中自体脂肪移植技术因脂肪组织来源广泛、获取途径相对简便、自体来源无免疫原性等优势已经成为近年来应用最广泛的组织移植技术, 在整形修复和美容医学领域均有重要的作用。但是,目前自体脂肪组织移植仍面临诸多挑战:移植物远期存留率波动较大,缺乏相应的干预和预测手段,限制了其进一步的临床开展和应用。
近年来,脂肪来源干细胞被证明对于脂肪移植后的存留具有重要的协同促再生作用。自体脂肪组织移植物中,脂肪细胞、脂肪来源千细胞、脂肪前体细胞、血管内皮细胞、上皮细胞、巨噬细胞等各个细胞组分所占比例各不相同,根据脂肪替代理论,移植物中非外周区域的脂肪细胞在移植术后,因缺氧缺血,发生大量的炎性坏死,而后脂肪来源干细胞通过增殖与分化作用逐渐替代原有的脂肪细胞区域;同时其旁分泌功能产生的炎症调节因子和活性细胞因子,可以调节受区的炎症反应,募集相关细胞向受区迁移,促进新生血管的形成和长入,进一步提供细胞增殖和分化所需要的氧气和营养物质,逐渐完成最终的替代过程,形成稳定的移植物容积。因此保留或增加脂肪来源干细胞的活性,同时抑制脂肪来源干细胞的衰老和死亡,可能有利于替代过程的发生,从而进一步增加脂肪移植物的远期存留率。
为了提高移植脂肪细胞的成活率,人们尝试在脂肪细胞中加入其他成分,例如Shin等在自体脂肪隆胸的脂肪细胞中加入ADSCs,术后都得到了较高的脂肪体积保留率,而且通过对比发现,加入的ADSCs量越大,效果越好。在脂肪移植时加入PRP可以增强ADSCs增殖和分化能力,促进移植后脂肪的血管化,改善局部血液循环,从而提高脂肪的成活率。Li等利用雷帕霉素的促自噬作用缓解ADSCs的凋亡,促进其分化成熟,从而提高移植后脂肪成活率。Kim等在动物实验中通过在移植脂肪中加入胰岛素与其他方式做对比,认为胰岛素可以促进ADSCs分化和存活。Wu等给裸鼠注射肉毒素来减少肌肉活动,然后再进行脂肪移植,发现脂肪细胞成活率明显提高。其他还有诸如在脂肪移植时应用透明质酸酶、雌二醇、热休克蛋白、三七皂苷等来提高脂肪细胞成活的实验,这些研究多为动物实验,其在临床应用效果尚难确定。
目前对于内皮细胞培养物对于移植脂肪的研究较少,Zhang等利用ADSCs和血管内皮细胞促进移植脂肪存活率,但效果有限。CN110229785A利用血管内皮祖细胞代替脂肪干细胞,能够有效提升脂肪移植后脂肪细胞的存活率。但现有技术中未有血管内皮培养上清液对移植脂肪存活率的研究。如何低成本地实现移植后脂肪细胞较高的存活率仍是当前研究的热点。
发明内容
基于此,有鉴于现有技术的上述缺陷,本发明所要解决的技术问题是提供一种成活率高的自体脂肪移植方法。
技术方案:为实现上述目的,我们提出了一种利用血管内皮细胞的上清添加到脂肪细胞和脂肪干细胞的混合物中脂肪细胞存活率,提高了脂肪体积保留率。
在本发明的一个实施方案中,提供了一种成活率高的自体脂肪移植方法,其包括以下步骤:
1)提取脂肪干细胞;
2)用血管内皮细胞培养得到的上清液处理脂肪干细胞;
3)将步骤2)中经处理的脂肪干细胞与颗粒脂肪混合,进行脂肪移植。
进一步地,所述上清液是血管内皮细胞先在常规条件下培养一段时间,然后在胁迫状态下培养一段时间后得到。
进一步地,所述胁迫状态是低血清、低氧和低一氧化氮的条件。
进一步地,所述血管内皮细胞是CRL-1730。
进一步地,所述上清液具体的制备步骤为:将CRL-1730常规培养于含10%胎牛血清的RPMI-1640培养液中培养1d,调整培养条件:1%胎牛血清,氧气5%,且培养液中加入40ml浓度为10mol/L的NO供体,培养6h,消化、离心、取上清液。
进一步地,所述脂肪干细胞为P3代。
进一步地,所述脂肪干细胞与颗粒脂肪混合的比例为1:19。
在本发明另一较佳的实施方案中,提供了一种血管内皮细胞培养得到的上清液在制备提高脂肪移植成活率的制剂中的应用。
进一步地,所述血管内皮细胞培养得到的上清液是是血管内皮细胞先在常规条件下培养一段时间,然后在胁迫状态下培养一段时间后得到。
进一步地,所述胁迫状态是低血清、低氧和低一氧化氮的条件。
进一步地,所述血管内皮细胞是CRL-1730。
有益效果:与现有技术相比,本发明制备的胁迫状态下内皮细胞上清液对于OGD条件下的脂肪干细胞的活性具有良好的保持作用;可以提高移植物最终的体积和质量保留率;可以抑制脂肪移植物凋亡;显著提高移植物新血管生成的能力;促进脂肪移植物的成活率。
以下将结合附图对本发明的构思、具体结构及产生的技术效果作进一步说明,以充分地了解本发明的目的、特征和效果。
附图说明
图1是本发明提取得到的脂肪干细胞的流式细胞术检测细胞表面标志物CD44、CD73、CD90、CD105、CD45的结果,图1A-D为CD44、CD73、CD90和CD105阳性表达,图1E为CD45阴性表达;
图2是脂肪干细胞在常规培养状态和OGD培养状态下经内皮细胞上清液预处理(ES)、胁迫状态下内皮细胞上清液组预处理(DES)以及等体积生理盐水处理(Sal)后的细胞活性图,图2A为常规培养状态,图2B为OGD培养状态;
图3是本发明移植物1m和3m的体积及质量保留率结果,图3A为体积保留率,图3B为质量保留率;
图4是体内实验中本发明移植物在1m和3mBAX基因的RT-PCR结果;
图5是本发明移植物在1m和3m的Caspase-3蛋白的Western Blot及灰度分析结果,图5A为Western Blot结果,图5B为灰度分析结果;
图6是体内实验中本发明移植物在1m和3mVEGFA基因的RT-PCR结果。
具体实施方式
以下参考说明书附图介绍本发明的多个优选实施例,使其技术内容更加清楚和便于理解。本发明可以通过许多不同形式的实施例来得以体现,本发明的保护范围并非仅限于文中提到的实施例。
脂肪细胞均来自整形外科医院腹部脂肪抽吸术的女性患者的术后废弃脂肪。纳入标准为:22-28岁,无既往病史,近3-6月无药物服用史。脂肪的采取及使用均征得患者本人同意。
实施例1
1、提取脂肪干细胞
将获得的脂肪剪碎,用PBS清洗3次,以100目的细胞筛过滤后,加入I型胶原酶消化,消化条件为:37℃恒温摇床消化60min。消化完成后,用DMEM培养基终止消化。以1000r的速度离心10分钟后用PBS清洗,重复两次。重悬后,再次低速离心10分钟。收集离心管底部细胞团即基质血管成分(SVF,Stromal Vascular Fraction), 用2ml的MSCM完全培养基重悬后以1×105/ml的密度接种于10cm细胞培养皿中,置于5%CO2、37℃条件下培养。原代细胞接种48小时后更换新培养基, 其后每48小时更换培养基。细胞达到90%融合后传代扩增,后续实验使用传至第三代(P3)的细胞。
2、脂肪干细胞多向分化能力的鉴定
P3细胞采用胰酶消化获取,以1×106个细胞/ml的浓度分离、洗涤和再悬浮细胞。抗CD44、CD73、CD90、CD105、CD45的抗体顺序添加到细胞悬浮液中,避光培30分钟,之后将细胞洗涤两次,在BD流式细胞仪上机检测。
3、多向分化潜能
分别将P3细胞在成脂(当细胞达到100%汇合时,每3天用含有10%FBS、1%抗生素/抗真菌溶液、0.5mM异丁基甲基黄嘌呤、1μM地塞米松、10μM胰岛素和200μM吲哚美辛的DMEM替换培养基,然后用含有10%FBS、1%抗生素/抗真菌剂的DMEM替换,在37℃、5%CO2条件下,每天摄入10μM胰岛素培养21天)、成骨(在含有10%FBS、1%抗生素/抗真菌剂、0.01μM 1,25-二羟基维生素D3、50μM抗坏血酸-2-磷酸盐和10mMβ-甘油磷酸盐的DMEM中培养。培养基每3天更换一次。在37℃、5%CO2条件下培养21天)和成软骨(将5×105ADSCs加入15ml试管中,用DMEM洗涤两次,并在200g下离心5min。沉淀物悬浮在含有10%FBS、1%抗生素/抗真菌剂、10%地塞米松、1%转化生长因子β3(TGF-β3)和1%其补充剂的DMEM中,每2-3天更换一次培养基。在37℃、5%CO2条件下培养28天)条件下诱导,观察其成脂、成骨和成软骨的能力。
结果表明,流式细胞结果如图1所示,待测细胞表面标志物CD44、CD73、CD90和CD105呈阳性表达,CD45呈阴性表达。
经诱导分化后,分别用油红O、茜素红和阿利新蓝染色,分别观察到艳红色成熟圆形脂滴、深红色钙质沉积和匀质蓝色软骨纤维,说明脂肪干细胞成功诱导为脂肪细胞、骨细胞和软骨细胞。
以上结果表明实施例1成功提取得到脂肪干细胞。
实施例2
人脐静脉血管内皮细胞CRL-1730购买自ATCC细胞库。
内皮细胞组:将CRL-1730常规培养于含10%胎牛血清的RPMI-1640培养液中,3d后,消化备用。
内皮细胞上清液组:将CRL-1730常规培养于含10%胎牛血清的RPMI-1640培养液中,3d后,消化、离心、取上清液备用。
胁迫状态下内皮细胞上清液组:将CRL-1730常规培养于含10%胎牛血清的RPMI-1640培养液中培养1d,调整培养条件:1%胎牛血清,氧气5%,且培养液中加入40ml浓度为10mol/L的NO供体,培养6h,消化、离心、取上清液备用。
实施例3
选取6周龄雌性BALB/c-nu/nu无胸腺小鼠(裸鼠)作为本部分研究的实验动物,裸鼠为SPF级并于IVC室中严格饲养,裸鼠购置于北京维通利华实验动物技术有限公司。
1、获取颗粒脂肪
将腹部脂肪抽吸术抽取的废弃脂肪,静置分层后去除水分,然后1200g离心3min,再次去除水分,手术剪将脂肪仔细修剪至直径约1mm的颗粒脂肪,备用。
将实验分为4组:脂肪干细胞与内皮细胞以5:1比例共培养3d的内皮细胞组(EC)、内皮细胞上清液预处理的内皮上清组(ES)、胁迫状态下内皮细胞上清液组预处理的胁迫上清组(DES)以及等体积生理盐水处理的脂肪干细胞对照组(Sal)。每组脂肪干细胞悬液与颗粒脂肪以1:19的体积比进行混合,充分混合后置于1ml的注射器中备用。
细胞活性
将内皮细胞上清液预处理的内皮上清组、胁迫状态下内皮细胞上清液组预处理的胁迫上清组以及等体积生理盐水处理的脂肪干细胞对照组分别在常规培养条件和缺氧乏糖(OGD)条件下培养1d,用cck-8检测脂肪干细胞活性。
3、动物模型
选择6周龄雌性裸鼠,在其背部选取4个位点,分别注射不同组的脂肪移植物,1m和3m后处死小鼠,进行取材。
4、移植物体积及质量保留率的测定
分别采用液体溢出法和精密电子天平检测移植物在移植前、后的体积与质量并计算体积和质量保留率。
5、实时荧光定量PCR检测
每100mg的移植物组织加1ml的TRlzol试剂,并用匀浆器匀浆至无颗粒透明状;将组织裂解液转移至离心管中,在室温下放置5min,使得核酸蛋白复合物完全分离后以1200g的速度在4℃离心10min,小心吸取上清液,移入新的离心管中;从向裂解液中加入氯仿开始步骤与第二部分相同,最终使用NanoDrop 2000c仪器测定RNA的浓度及纯度。
各基因所用的双向引物序列见下表:
6、Western Blot检测
称量组织块后用玻璃匀浆器研磨组织,10ml预冷的PBS洗一遍。将匀浆转移到离心管中,以1500r/min的速度在4℃下离心5min;预冷裂解液,临用前以100:1的体积比加入终浓度为1x的PMSF蛋白酶抑制剂。随后加入裂解液(每1mg组织加入约4μL的裂解液),吹打混匀,放置冰上。每隔5-6分钟轻柔振荡一次,共4次;以1000g的速度在4℃下离心3min后取上清。进行蛋白浓度测定及后续的Western Blot检测。
结果表明,cck-8细胞活性检测如图2所示,通过图2A和图2B的对比,可知胁迫状态下内皮细胞上清液对于OGD条件下的脂肪干细胞的活性具有良好的保持作用;移植1m和3m的体积及质量保留率均表明,DES组体积保留率和质量保留率明显高于其他三组,说明胁迫状态下内皮细胞上清液可以提高移植物最终的体积和质量保留率;对于移植1m和3m后移植物的凋亡水平,移植1m和3m的DES组的BAX基因表达水平明显低于其他三组,说明胁迫状态下内皮细胞上清液能抑制脂肪移植物凋亡,同样在蛋白水平,凋亡蛋白Caspase-3的WB结果同样证明上述结论;对于移植1m和3m后移植物的新血管生成情况,通过VEGFA基因的表达水平来评价,其RT-PCR结果显示,DES组的VEGFA基因表达水平明显高于其他三组,可以推断是胁迫状态下内皮细胞上清液显著提高了移植物新血管生成的能力,进而促进了脂肪移植物的成活率。
以上详细描述了本发明的较佳具体实施例。应当理解,本领域的普通技术无需创造性劳动就可以根据本发明的构思作出诸多修改和变化。因此,凡本技术领域中技术人员依本发明的构思在现有技术的基础上通过逻辑分析、推理或者有限的实验可以得到的技术方案,皆应在由权利要求书所确定的保护范围内。
Claims (10)
1.一种成活率高的自体脂肪移植方法,其包括以下步骤:
1)提取脂肪干细胞;
2)用血管内皮细胞培养得到的上清液处理脂肪干细胞;
3)将步骤2)中经处理的脂肪干细胞与颗粒脂肪混合,进行脂肪移植。
2.根据权利要求1所述的自体脂肪移植方法,其特征在于,所述上清液是血管内皮细胞先在常规条件下培养一段时间,然后在胁迫状态下培养一段时间后得到的。
3.根据权利要求2所述的自体脂肪移植方法,其特征在于,所述胁迫状态是低血清、低氧和低一氧化氮的条件。
4.根据权利要求1所述的自体脂肪移植方法,其特征在于,所述血管内皮细胞是CRL-1730。
5.根据权利要求3所述的自体脂肪移植方法,其特征在于,所述上清液具体的制备步骤为:将CRL-1730常规培养于含10%胎牛血清的RPMI-1640培养液中培养1d,调整培养条件:1%胎牛血清,氧气5%,且培养液中加入40ml浓度为10mol/L的NO供体,培养6h,消化、离心、取上清液。
6.根据权利要求1所述的自体脂肪移植方法,其特征在于,所述脂肪干细胞为P3代。
7.根据权利要求1所述的自体脂肪移植方法,其特征在于,所述脂肪干细胞与颗粒脂肪混合的比例为1:19。
8.血管内皮细胞培养得到的上清液在制备提高脂肪移植成活率的制剂中的应用。
9.根据权利要求8所述的应用,其特征在于,所述血管内皮细胞培养得到的上清液是是血管内皮细胞先在常规条件下培养一段时间,然后在胁迫状态下培养一段时间后得到。
10.根据权利要求9所述的应用,其特征在于,所述胁迫状态是低血清、低氧和低一氧化氮的条件。
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Citations (13)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20070202592A1 (en) * | 2004-07-08 | 2007-08-30 | Yasuo Kitagawa | Pluripotent Cells Distributed Ubiquitously In Animal Tissue, Which Proliferate Selectively In Lower-Serum Culture |
KR20100007113A (ko) * | 2008-07-11 | 2010-01-22 | 김민 | 광 처리된 지방조직 세포 유래 이식용 조성물 |
KR20150049041A (ko) * | 2013-10-29 | 2015-05-08 | 주식회사 엔바이오텍 | 지방유래 줄기세포 배양방법 |
CN106421920A (zh) * | 2016-11-16 | 2017-02-22 | 沈阳细胞治疗工程技术研发中心有限公司 | 一种脂肪填充物及其制备方法 |
CN106701668A (zh) * | 2015-07-22 | 2017-05-24 | 中国医药大学 | 间质干细胞、其纯株化扩张的方法、其分离方法及其应用 |
CN108624557A (zh) * | 2018-05-31 | 2018-10-09 | 章毅 | 间充质干细胞外泌体的制备方法及其应用 |
CN110229785A (zh) * | 2019-06-17 | 2019-09-13 | 白晋 | 一种利用血管内皮祖细胞提升脂肪移植成功率的方法 |
CN111228571A (zh) * | 2020-03-25 | 2020-06-05 | 白晋 | 一种注射用丰唇填充物的制备方法 |
CN111304150A (zh) * | 2020-02-25 | 2020-06-19 | 昆明医科大学 | 可促进移植物微循环血供恢复的ADSCs与EPCs干细胞体系 |
CN112111451A (zh) * | 2020-11-23 | 2020-12-22 | 北京欣颂生物科技有限公司 | 提高干细胞细胞因子产量的方法 |
CN114736853A (zh) * | 2022-04-02 | 2022-07-12 | 麦迪森(江苏)医学研究有限公司 | 一种增强间充质干细胞促血管新生能力的方法 |
CN115725500A (zh) * | 2022-12-01 | 2023-03-03 | 徐州医科大学附属医院 | 脂肪间充质干细胞外泌体、其制备方法及其应用 |
US20230174941A1 (en) * | 2020-07-17 | 2023-06-08 | Medical Corporation Yanaga Clinic | Method for producing mature adipocyte-containing composition |
Patent Citations (13)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20070202592A1 (en) * | 2004-07-08 | 2007-08-30 | Yasuo Kitagawa | Pluripotent Cells Distributed Ubiquitously In Animal Tissue, Which Proliferate Selectively In Lower-Serum Culture |
KR20100007113A (ko) * | 2008-07-11 | 2010-01-22 | 김민 | 광 처리된 지방조직 세포 유래 이식용 조성물 |
KR20150049041A (ko) * | 2013-10-29 | 2015-05-08 | 주식회사 엔바이오텍 | 지방유래 줄기세포 배양방법 |
CN106701668A (zh) * | 2015-07-22 | 2017-05-24 | 中国医药大学 | 间质干细胞、其纯株化扩张的方法、其分离方法及其应用 |
CN106421920A (zh) * | 2016-11-16 | 2017-02-22 | 沈阳细胞治疗工程技术研发中心有限公司 | 一种脂肪填充物及其制备方法 |
CN108624557A (zh) * | 2018-05-31 | 2018-10-09 | 章毅 | 间充质干细胞外泌体的制备方法及其应用 |
CN110229785A (zh) * | 2019-06-17 | 2019-09-13 | 白晋 | 一种利用血管内皮祖细胞提升脂肪移植成功率的方法 |
CN111304150A (zh) * | 2020-02-25 | 2020-06-19 | 昆明医科大学 | 可促进移植物微循环血供恢复的ADSCs与EPCs干细胞体系 |
CN111228571A (zh) * | 2020-03-25 | 2020-06-05 | 白晋 | 一种注射用丰唇填充物的制备方法 |
US20230174941A1 (en) * | 2020-07-17 | 2023-06-08 | Medical Corporation Yanaga Clinic | Method for producing mature adipocyte-containing composition |
CN112111451A (zh) * | 2020-11-23 | 2020-12-22 | 北京欣颂生物科技有限公司 | 提高干细胞细胞因子产量的方法 |
CN114736853A (zh) * | 2022-04-02 | 2022-07-12 | 麦迪森(江苏)医学研究有限公司 | 一种增强间充质干细胞促血管新生能力的方法 |
CN115725500A (zh) * | 2022-12-01 | 2023-03-03 | 徐州医科大学附属医院 | 脂肪间充质干细胞外泌体、其制备方法及其应用 |
Non-Patent Citations (6)
Title |
---|
刘林奇: ""低氧预处理的人脂肪来源干细胞促进血管生成的体外实验研究"", 《南方医科大学硕士学位论文》, 15 April 2014 (2014-04-15), pages 15 - 32 * |
李明等: ""血管内皮细胞上清液对口腔颊黏膜成纤维细胞增殖活性的影响"", 《临床口腔医学杂志》, vol. 26, no. 2, 26 April 2010 (2010-04-26), pages 89 - 91 * |
杨艳清;许其军;张洁;吴希;黄燕;江昌艳;: "不同部位来源脂肪干细胞促进血管化的作用比较", 华中科技大学学报(医学版), no. 03, 15 June 2018 (2018-06-15) * |
钟晓春;倪有娣;何晓升;: "脂肪来源的间充质干细胞对颗粒状脂肪组织移植效果的影响", 健康研究, no. 06, 15 December 2009 (2009-12-15) * |
闫静川, 脂肪干细胞培养上清液对体外培养人脐静脉血管内皮细胞的影响, no. 2021, 31 May 2021 (2021-05-31), pages 211 - 214 * |
陈临溪: "《血管内皮细胞药理与临床》", 31 December 2012, 人民军医出版社, pages: 11 - 12 * |
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