CN108300686B - 用于预防、延缓或逆转细胞、组织、器官、机体衰老的小分子化合物/组合,产品及其用途 - Google Patents
用于预防、延缓或逆转细胞、组织、器官、机体衰老的小分子化合物/组合,产品及其用途 Download PDFInfo
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Abstract
本发明公开了一种用于预防、延缓或逆转细胞、组织、器官、机体衰老的小分子化合物/组合,产品及其用途。所述小分子化合物组合包括DNMT抑制剂、HMT抑制剂、组蛋白去甲基化酶抑制剂、TGF‑β抑制剂,WNT/β‑catenin激动剂、cAMP激动剂、赖氨酸脱乙酰基酶抑制剂、RAR激动剂和ROCK抑制剂中的至少一种。本发明采用小分子化合物或其组合预防、延缓或逆转细胞、组织、器官、机体衰老,延长细胞端粒长度,作用靶点明确、成份清楚、成熟稳定、可准确调控抗衰老的进程;并且小分子化合物易于质控及规模化生产。由于在作用过程中机体或细胞不表达干性基因,安全性高;而且小分子化合物及其组合对机体易于吸收,能够分别在细胞、组织、器官、机体的整体水平上,预防、延缓和逆转老化,对延缓人类衰老进程、提高健康状态及生活质量具有重要意义。
Description
技术领域
本发明涉及细胞生物学,组织工程和医学,尤其涉及一种用于预防、延缓或逆转细胞、组织、器官、机体衰老的小分子化合物/组合,产品及其用途。
背景技术
人类机体衰老是现代社会的主要健康问题,也是大部分老年性疾病的主要原因。衰老过程伴随身体机能的降低、对外界变化适应能力的减弱、代偿功能的低下,最终诱发多种疾病。如心血管疾病、恶性肿瘤、神经、内脏、骨骼和肌肉的退行性疾病、糖尿病等都与衰老密切相关。年龄每增加10岁,因心血管病的病死率即增加2~3倍,可见衰老对人类生活质量与健康状态影响巨大。
近几十年来,与衰老相关的因素(如衰老相关基因)以及抗衰老药物、方法不断被报道。例如2016年,美国梅奥诊所的研究团队通过清除衰老细胞,使小鼠的寿命延长超过20%;不同的研究机构发现通过喂食小鼠雷帕霉素、辅酶I、NMN(烟酰胺单核苷酸)、亚精胺等可以显著延长小鼠的寿命;有研究机构利用线虫模型揭示了RNA剪接(RNA splicing)功能与长寿之间的关联;另有研究机构报道了HOXA9在老年时的重激活会限制骨骼肌的再生,NRF2的激活有利于抗衰老;另外,据有关报道,清除线粒体中突变的DNA,或调整跑步、节食等生活习惯与生活方式对延缓衰老也有积极作用。
尽管上述的物质或者方法都有改善器官健康状态,从而提高寿命的作用,但鉴于细胞、组织、器官水平衰老进程的复杂性,人们对衰老进程及其机制的了解仍然有限,上述各种延缓衰老的方法和物质的作用效果仍具有不确定性。2016年12月,索尔克研究所的研究人员通过在活体小鼠中高表达Yamanaka四因子(制备诱导多能性干细胞的四因子)的方式,让早衰症小鼠的全身很多器官恢复到相对健康年轻的状态,并延长了小鼠1/3的寿命。因此,用于延缓或者逆转机体大部分细胞或组织器官衰老进程的方法或者物质,如果作用明确,过程可控,有望成为人类抗衰老的可靠技术基础。
转基因技术虽然在早衰症小鼠身上实现了全身大部分脏器的衰老逆转,但鉴于安全性考虑,此方法在人体内不可行。此外,使用的Yamanaka四因子的激活有致瘤的风险。因此,有必要寻找可用于人体的安全有效的作用方式和物质。小分子化合物或者小分子化合物组合因其具备以下优势:①不导入外源性转录因子,不改变源细胞的基因结构,具有良好的安全性与稳定性;②作用***较为稳定,易于质控,成本低廉;③作用过程短,效率高,便于规模化生产;④便于人体吸收和代谢,将成为减缓或者逆转衰老进程的有效物质。然而,小分子化合物或者小分子化合物组合抗衰老的的报道和研究很少,一方面由于缺乏筛选该类小分子化合物的方法和体系;另一方面,即使在小鼠上尝试有用的小分子化合物,由于人类与小鼠存在约25%的基因差异,应用于人的同类细胞,其可行性也不高。因此,有针对性地筛选可安全并明确地延缓或逆转人类衰老进程的小分子化合物,对延缓人类衰老进程、提高健康状态及生活质量具有重要意义。
发明内容
本发明提供一种用于预防、延缓或逆转细胞、组织、器官、机体衰老的小分子化合物/组合,产品及其用途。所述小分子化合物组合在细胞、组织、器官、机体抗衰老的过程中具有可调控性、整体性、可逆转性和安全性。
本发明第一方面,提供一种预防、延缓或者逆转哺乳动物如人的细胞、组织、器官或机体衰老进程的小分子化合物组合,所述小分子化合物组合包括DNMT抑制剂、HMT抑制剂、组蛋白去甲基化酶抑制剂、TGF-β抑制剂、WNT/β-catenin激动剂、cAMP激动剂和赖氨酸脱乙酰基酶抑制剂中的至少一种。
作为选择,所述小分子化合物组合为DNMT抑制剂、HMT抑制剂、组蛋白去甲基化酶抑制剂和赖氨酸脱乙酰基酶抑制剂中的至少一种。
作为选择,所述小分子化合物组合为TGF-β抑制剂、WNT/β-catenin激动剂、cAMP激动剂、DNMT抑制剂、HMT抑制剂和赖氨酸脱乙酰基酶抑制剂中的至少一种。
进一步地,所述小分子化合物还包括RAR激动剂、ascorbate(抗坏血酸)和ROCK抑制剂中的至少一种。
进一步地,所述小分子化合物组合包括按时序分阶段使用的第一阶段化合物和第二阶段化合物,所述第一阶段化合物为DNMT抑制剂、HMT抑制剂、组蛋白去甲基化酶抑制剂、TGF-β抑制剂、WNT/β-catenin激动剂、cAMP激动剂和赖氨酸脱乙酰基酶抑制剂中的至少一种;
所述第二阶段化合物为DNMT抑制剂、HMT抑制剂、组蛋白去甲基化酶抑制剂、TGF-β抑制剂,WNT/β-catenin激动剂、cAMP激动剂、赖氨酸脱乙酰基酶抑制剂、RAR激动剂、ascorbate(抗坏血酸)和ROCK抑制剂中的至少一种。
所述TGF-β受体抑制剂为I型TGF-β受体抑制剂,所述cAMP激动剂为EPAC/RAP1激动剂。
进一步地,所述小分子化合物组合与PDGF-AB组合使用,提高小分子化合物组合的作用效率。
作为优选,所述赖氨酸脱乙酰基酶抑制剂包括sodium phenylbutyrate,butyrate,sodium butyrate,MC1568,CI994(Tacedinaline),chidamide,CAY10683(SantacruzaMate A),CUDC-907,M344(Histone Deacetylase Inhibitor III),LAQ824(NVP-LAQ824,Dacinostat),Prac inostat(SB939),VPA(Valproic acid),Valproic acidsodium salt,Scriptaid,Apicidin,LBH-589(Panobinostat),MS-275,SAHA(Vorinostat),Trichostatin(TSA),Psammaplin A,PCI-24781(A bexinostat),Rocilinostat(ACY-1215),Mocetinostat(MGCD0103),4-Phenylbutyrate(4PB),splitomicin,SRT1720,resveratrol,Sirtinol,APHA,CI-994,Depudecin,FK-228,HC-Toxin,ITF-2357(Givinostat),Chidamide,RGFP 966,PHOB,BG45,Nexturastat A,TMP269,CAY10603,MGCD-0103,Niltubacin,PXD-101(Belinostat),Pyroxamide,Tubacin,EX-527,BATCP,Cambinol,MOCPAC,PTACH,MC1568,NCH51和TC-H106中的至少一种;
所述TGF-β受体抑制剂包括616452,LY2109761,Pirfenidone,Repsox(E-616452),SB431542,A77-01,Tranilast,Galunisertib(LY2157299),A8301,GW788388,ITD-1,SD208,SB525334,LY364947,ASP3029,D4476和SB505124中的至少一种;
所述WNT/β-catenin激动剂包括MAY-262611,CHIR98014,CHIR99021,LiCl,Li2CO3,TD114-2,AZD2858,AZD1080,BIO,Kenpaullone,TWS119,LY2090314,CBM1078,SB216763和AR-A014418中的至少一种;
所述cAMP激动剂包括EPAC/RAP1激动剂,8-Bromo-cAMP,Dibutyryl-Camp和Sp-8-Br-cAMPs中的至少一种;
所述EPAC/RAP1激动剂包括Forskolin,IBMX,Prostaglandin E2(PGE2),NKH477,8-pCPT-2′-O-Me-cAMP,GSK256066,Apremilast(CC-10004),Roflumilast,Cilomilast,Rolipram和Milrinone中的至少一种;
所述RAR激动剂包括TTNPB,Bexarotene,Ch55,Tamibarotene,Retinol,AM580,ATRA,13-cis RA,Vitamin A及Vitamin A衍生物中的至少一种;
所述ROCK抑制剂包括Y-27632,Y-27632 2HCl,Thiazovivin,Ripasudil(K-115),Fasudil,Fasudil(HA-1077)HCl,GSK429286A,RKI-1447和PKI-1313中的至少一种;
所述DNMT抑制剂包括RG108,Thioguanine,5-Aza-2′-deoxycytidine(Decitabine),SGI-1027,Zebularine和5-Azacytidine(AZA)中的至少一种;
所述HMT抑制剂包括EPZ004777,EPZ5676,GSK503,BIX 01294,DZNep,DZNep HCL,SGC 0946中的至少一种;
所述组蛋白去甲基化酶抑制剂包括parnate(tranylcypromine),Tranylcypromine(2-PCPA)HCl SP2509,4SC-202,ORY-1001(RG-6016),GSKJ1和GSK-LSD1中的至少一种。
作为进一步优选,所述赖氨酸脱乙酰基酶抑制剂为sodium butyrate,VPA和Trichostatin(TSA)的至少一种;
所述TGF-β受体抑制剂为Repsox(E-616452)和SB431542中的至少一种;
所述WNT/β-catenin激动剂为CHIR99021和BIO中的至少一种;
所述cAMP激动剂为Forskolin和Rolipram中的至少一种;
所述RAR激动剂为TTNPB和AM580中的至少一种;
所述ROCK抑制剂包括Y-27632和Thiazovivin中的至少一种;
所述DNMT抑制剂为RG108,5-Aza-2′-deoxycytidine(Decitabine)和5-Azacytidine(AZA)中的至少一种;
所述HMT抑制剂为EPZ004777,EPZ5676和SGC 0946中的至少一种;
所述组蛋白去甲基化酶的抑制剂为parnate(tranylcypromine)和Tranylcypromine(2-PCPA)HCl的至少一种。
所述小分子化合物组合用于延长哺乳动物如人的细胞的端粒长度,瞬时刺激端粒相关蛋白的表达或上调,如TERT(端粒酶反转录酶)的表达和TPP1上调;作用后不会刺激哺乳动物如人的细胞表达干性基因,如OCT4,Nanog等。
本发明的一些实施方案,具体小分子化合物的有效浓度如下,以下给出的浓度范围只是参考,可在此基础上做适应性修改,如果其他小分子替代以下小分子,浓度也可以做适应性调整。
Forskolin浓度为2μM~20μM;Repsox浓度为2~15uM;CHIR99021浓度为1μM~10μM;VPA浓度为0.5mM~1.5mM;TTNPB浓度为3μM~8μM;AM580浓度为0.03~0.08μM;EPZ004777浓度为3~8μM;Y-27632浓度为3~15μM,L-Ascorbin acid 2-phosphate浓度为0.15~0.25mM;5-Aza-2′-deoxycytidine浓度为0.5~20μM。
本发明预防、延缓或逆转衰老的机理如下:本发明通过对老化或者衰老的细胞进行表观遗传学上的调节,或者通过激活早期发育相关的转录因子,如激活WNT/β-catenin信号通路、激活cAMP信号通路、激动RA信号通路,抑制TGF-beta信号通路,实现细胞衰老的预防,延缓或者逆转。所述小分子化合物组合可用于延长哺乳动物如人的细胞的端粒长度,瞬时刺激端粒相关蛋白的表达/上调,如TERT(端粒酶反转录酶)和TPP1,不会刺激哺乳动物如人的细胞表达干性基因,如OCT4,Nanog等,安全有效。
本发明第二方面,提供一种包含所述小分子化合物组合的试剂盒、培养液/基、药物、保健品、食品、化妆品或医疗器械。
本发明第三方面,提供一种所述小分子化合物组合在预防、延缓或者逆转哺乳动物如人的细胞、组织、器官或机体衰老进程和相关疾病中的应用及提高哺乳动物如人组织、器官修复能力的应用。
所述细胞来源于哺乳动物如人的机体细胞或体外培养的细胞。
本发明第四方面,提供一种由所述小分子化合物组合预防、延缓或者逆转哺乳动物如人的细胞、组织、器官或机体衰老进程的方法。
本发明第五方面,提供一种通过所述小分子化合物组合作用得到的年轻化的细胞及细胞产物。
本发明第六方面,提供一种所述年轻化的细胞及细胞产物的应用。
本说明书中采用的术语“衰老”与“老化”的意思相同,年轻化的细胞指的是衰老前的细胞。但本发明中所述的“衰老”不包含永久停止***的细胞。
本发明具有以下有益效果:本发明采用小分子化合物或其组合预防、延缓或逆转细胞、组织、器官、机体衰老,延长细胞端粒长度,作用靶点明确、成份清楚、成熟稳定、可准确调控抗衰老的进程;并且小分子化合物易于质控及规模化生产。由于在作用过程中机体或细胞不表达干性基因,安全性高;而且小分子化合物及其组合对机体易于吸收,能够分别在细胞、组织、器官、机体的整体水平上,预防、延缓和逆转老化,对延缓人类衰老进程、提高健康状态及生活质量具有重要意义。
附图说明
图1为小分子化合物作用后人源皮肤成纤维细胞形态图;
图2为小分子化合物作用后人源骨髓间充质干细胞形态图;
图3为小分子化合物作用后人源皮肤成纤维细胞细胞性状的鉴定;
图4为小分子化合物作用后人源骨髓间充质干细胞细胞性状的鉴定;
图5为通过小分子化合物作用后人源细胞的端粒长度变化;
图6为通过小分子化合物作用后人源细胞的端粒相关基因的表达;
图7为对小分子化合物作用后人源皮肤成纤维细胞中与衰老相关基因的表达变化图;
图8为小分子化合物作用后人源皮肤成纤维细胞体外功能活力的检测结果图;
图9为小分子化合物作用后人源皮肤成纤维细胞体内功能活力的检测结果图;
图10为对小分子化合物作用后长期传代的细胞干性的检测和成瘤性检测结果图;
图11为长期传代的小分子化合物作用后人源皮肤成纤维细胞的克隆形成及形态图。
具体实施方式
下面结合附图和具体实施例对本发明的技术方案做进一步详细说明,但本发明并不局限于以下技术方案。
实施例1
1、皮肤成纤维细胞的分离
1.1从志愿者身上获取直径约1cm的皮肤组织块,贴壁法分离皮肤成纤维细胞,分离的细胞培养于基础培养液里,所述基础培养液:10%胎牛血清(Hyclone)+100U/ml青霉素(Sigma)+100μg/ml链霉素(Sigma)+高糖DMEM。
1.2细胞传代大量扩增,细胞代数在本实验中,细胞为第12代,处理的前一天(Day-1),接种细胞密度1~2.5×104/cm2培养于37℃,5%CO2的培养箱中。
2、皮肤成纤维细胞的小分子化合物处理
更换基础培养液为小分子处理液,培养细胞2-20天,小分子处理液是指:10%胎牛血清(Hyclone)+100U/ml青霉素(Sigma)+100μg/ml链霉素(Sigma)+高糖DMEM培养基(Gibco)+forskolin(2μM~25μM)+Repsox(2~15uM)+CHIR99021(1μM~10μM)+VPA(0.5mM~1.5mM)+5-Aza-2′-deoxycytidine(0.5~20μM),该培养***中10%胎牛血清也可以被血清替代品(invitrogen)以10%~20%的浓度替代;100U/ml青霉素(Sigma)和100μg/ml链霉素(Sigma)可以不使用。在37℃,5%CO2环境下培养细胞。
3、皮肤成纤维细胞的维持培养
上述第2步骤的处理结束后,完全更换为基础培养液,正常维持培养,在37℃,5%CO2环境下培养细胞。所述基础培养液:10%胎牛血清(Hyclone)+100U/ml青霉素(Sigma)+100μg/ml链霉素(Sigma)+高糖DMEM,中10%胎牛血清也可以被血清替代品(invitrogen)以10%~20%的浓度替代;100U/ml青霉素(Sigma)和100μg/ml链霉素(Sigma)可以不使用。
4、处理后细胞性状的鉴定
4.1细胞传代到第14代时,使用碧云天细胞衰老β-半乳糖苷酶染色试剂盒进行染色鉴定;
4.2细胞传代到第14代,使用ebioscience的CD29-PE流式抗体进行染色,对照为同一厂家对应的同型对照品,按照厂家说明书操作,在BD JAZZ流式仪上进行分析;
4.3细胞传代到第14代,在24孔板里培养,使用4%的PFA固定细胞10分钟后,按照免疫荧光的操作步骤,使用santa cruz的Vimentin抗体进行染色;
4.4细胞增值的检测,以1.5x105作为起始接种细胞量,连续12天计数细胞,每个时间点重复3组;
4.5细胞传代到第14代,收取DNA样本,使用Cawthon qPCR方法进行端粒长度的检测,荧光染料采用Takara SYBR Green I,
端粒扩增引物序列为
mTel:CGGTTTGTTTGGGTTTGGGTTTGGGTTTGGGTTTGGGTT;
mTel:GGCTTGCCTTACCCTTACCCTTACCCTTACCCTTACCCT;
单拷贝基因引物
36B4-S-F:CAGCAAGTGGGAAGGTGTAATCC;
36B4-S-R:CCCATTCTATCATCAACGGGTACAA;
4.6细胞传代到第14代,收取RNA样本,采用PCR方法进行TERT的检测,人源TERT的引物序列为F:5′-CGGAAGAGTGTCTGGAGCAA-3;R5′-GGATGAAGCGGAGTCTGGA-3′;
4.7细胞传代到第14代,收取RNA样本,通过转录组的分析,定量基因TPP1、CDKN1A、ATF3、GADD45B、BTG2、H2AFX、ITGA6、COL1A1表达量的变化;
4.8异体皮肤移植,检测小分子化合物处理后的人源皮肤成纤维细胞培养液的免疫调节作用。具体方法为:实验组为小分子化合物处理细胞组,未处理细胞组,PBS组,其中传代到第14代,按照动物伦理委员的要求执行动物实验,将供体C57B/6小鼠麻醉,仰卧位固定,全背部脱毛,背部取1.5cm x 1.5cm的全层的皮肤,去除皮下脂肪及血管,将小鼠皮肤剪成直径为1.0cm大小的皮片,置含有无菌PBS的平皿中备用。将剪好的皮片翻转过来放入有生理盐水的培养皿中,用手术刀片轻轻地切去皮下组织至真皮,然后放在无菌生理盐水中备用。
受体为BALB/c小鼠,麻醉后背部脱毛,碘酒消毒背部皮肤,制备一直径1.0cm的皮肤缺口,保留血管,于背部逆毛方向移植C57B/6皮片,表面敷盖凡士林油纱和创可贴,松紧适度。移植后1小时内,从尾静脉注射列表中的各组别液体。
4.9细胞克隆形成率的检测:细胞传代到第14代,取对数生长期的细胞,以10个细胞/cm2铺板,每组设3复孔,每隔2天换液,1周~2周左右Giemsa或者PI染色,显微镜下计数形成的克隆数量。
实施例2
1、骨髓间充质干细胞的培养
1.1购买商品化的人源骨髓间充质干细胞或者从骨创伤后志愿者骨碎片残留的骨髓中分离间充质干细胞,细胞培养于基础培养液里,所述基础培养液:8%胎牛血清(Hyclone)+100U/ml青霉素(Sigma)+100μg/ml链霉素(Sigma)+低糖DMEM。
1.2细胞传代大量扩增,细胞代数在本实验中,细胞为第10代,处理的前一天(Day-1),接种细胞密度1~2.5×104/cm2培养于37℃,5%CO2的培养箱中。
2、骨髓间充质干细胞的小分子化合物处理
更换基础培养液为小分子处理液,培养细胞2-20天,小分子处理液是指:10%胎牛血清(Hyclone)+100U/ml青霉素(Sigma)+100μg/ml链霉素(Sigma)+高糖DMEM培养基(Gibco)+forskolin(2μM~25μM)+Repsox(2~15uM)+CHIR99021(1μM~10μM)+VPA(0.5mM~1.5mM),该培养***中10%胎牛血清也可以被血清替代品(invitrogen)以10%~20%的浓度替代;100U/ml青霉素(Sigma)和100μg/ml链霉素(Sigma)可以不使用。在37℃,5%CO2环境下培养细胞。
3、骨髓间充质干细胞的维持培养
上述第2步骤的处理结束后,完全更换为基础培养液,正常维持培养,在37℃,5%CO2环境下培养细胞。所述基础培养液:8%胎牛血清(Hyclone)+100U/ml青霉素(Sigma)+100μg/ml链霉素(Sigma)+低糖DMEM,中10%胎牛血清也可以被血清替代品(invitrogen)以10%~20%的浓度替代;100U/ml青霉素(Sigma)和100μg/ml链霉素(Sigma)可以不使用。
4、处理后细胞性状的鉴定
4.1细胞传代到第12代时,使用碧云天细胞衰老β-半乳糖苷酶染色试剂盒进行染色鉴定;
4.2细胞传代到第12代,使用ebioscience的CD29-PE,CD90,CD73,CD34,CD45流式抗体进行染色,对照为同一厂家对应的同型对照品,按照厂家说明书操作,在BD JAZZ流式仪上进行分析;
4.3细胞传代到第12代,使用Cyagen Biosciences的成骨和成脂分化诱导液进行诱导,具体步骤按照产品说明书;
4.4细胞增值的检测,以1.5x105作为起始接种细胞量,连续12天计数细胞,每个时间点重复3组;
4.5细胞传代到第14代,收取DNA样本,使用Cawthon qPCR方法进行端粒长度的检测,荧光染料采用Takara SYBR Green I,
端粒扩增引物序列为
mTel:CGGTTTGTTTGGGTTTGGGTTTGGGTTTGGGTTTGGGTT;
mTel:GGCTTGCCTTACCCTTACCCTTACCCTTACCCTTACCCT;
单拷贝基因引物
36B4-S-F:CAGCAAGTGGGAAGGTGTAATCC;
36B4-S-R:CCCATTCTATCATCAACGGGTACAA;
4.6细胞传代到第14代,收取RNA样本,采用PCR方法进行TERT的检测,人源TERT的引物序列为F:5′-CGGAAGAGTGTCTGGAGCAA-3;R5′-GGATGAAGCGGAGTCTGGA-3′;
4.7细胞传代到第14代,收取RNA样本,通过转录组的分析,定量基因TPP1、CDKN1A、ATF3、GADD45B、BTG2、H2AFX表达量的变化;
4.8异体皮肤移植,检测小分子化合物处理后的人源骨髓间充质干细胞培养液的免疫调节作用。具体方法为:实验组为小分子化合物处理细胞组,未处理细胞组,PBS组,其中传代到第14代,按照动物伦理委员的要求执行动物实验,将供体C57B/6小鼠麻醉,仰卧位固定,全背部脱毛,背部取1.5cm x 1.5cm的全层的皮肤,去除皮下脂肪及血管,将小鼠皮肤剪成直径为1.0cm大小的皮片,置含有无菌PBS的平皿中备用。将剪好的皮片翻转过来放入有生理盐水的培养皿中,用手术刀片轻轻地切去皮下组织至真皮,然后放在无菌生理盐水中备用。
受体为BALB/c小鼠,麻醉后背部脱毛,碘酒消毒背部皮肤,制备一直径1.0cm的皮肤缺口,保留血管,于背部逆毛方向移植C57B/6皮片,表面敷盖凡士林油纱和创可贴,松紧适度。移植后1小时内,从尾静脉注射列表中的各组别液体。
4.9细胞克隆形成率的检测:细胞传代到第14代,取对数生长期的细胞,以10个细胞/cm2铺板,每组设3复孔,每隔2天换液,1周~2周左右Giemsa或者PI染色,显微镜下计数形成的克隆数量。
实施例3
1、皮肤成纤维细胞的分离
1.1从志愿者身上获取直径约1cm的皮肤组织块,贴壁法分离皮肤成纤维细胞,分离的细胞培养于基础培养液里,所述基础培养液:10%胎牛血清(Hyclone)+100U/ml青霉素(Sigma)+100μg/ml链霉素(Sigma)+高糖DMEM。
1.2细胞传代大量扩增,细胞代数在本实验中,细胞为第12代,处理的前一天(Day-1),接种细胞密度1~2.5×104/cm2培养于37℃,5%CO2的培养箱中。
2、皮肤成纤维细胞的小分子化合物处理
更换基础培养液为小分子处理液,培养细胞2-20天,小分子处理液是指:10%胎牛血清(Hyclone)+100U/ml青霉素(Sigma)+100μg/ml链霉素(Sigma)+高糖DMEM培养基(Gibco)+forskolin(2μM~25μM)+5-Aza-2′-deoxycytidine(0.5~20μM),该培养***中10%胎牛血清也可以被血清替代品(invitrogen)以10%~20%的浓度替代;100U/ml青霉素(Sigma)和100μg/ml链霉素(Sigma)可以不使用。在37℃,5%CO2环境下培养细胞。
3、皮肤成纤维细胞的维持培养
上述第2步骤的处理结束后,完全更换为基础培养液,正常维持培养,在37℃,5%CO2环境下培养细胞。所述基础培养液:10%胎牛血清(Hyclone)+100U/ml青霉素(Sigma)+100μg/ml链霉素(Sigma)+高糖DMEM,中10%胎牛血清也可以被血清替代品(invitrogen)以10%~20%的浓度替代;100U/ml青霉素(Sigma)和100μg/ml链霉素(Sigma)可以不使用。
4、处理后细胞性状的鉴定
详细步骤同实施例1
实施例4
1、皮肤成纤维细胞的分离
1.1从志愿者身上获取直径约1cm的皮肤组织块,贴壁法分离皮肤成纤维细胞,分离的细胞培养于基础培养液里,所述基础培养液:10%胎牛血清(Hyclone)+100U/ml青霉素(Sigma)+100μg/ml链霉素(Sigma)+高糖DMEM。
1.2细胞传代大量扩增,细胞代数在本实验中,细胞为第12代,处理的前一天(Day-1),接种细胞密度1~2.5×104/cm2培养于37℃,5%CO2的培养箱中。
2、皮肤成纤维细胞的小分子化合物处理
更换基础培养液为小分子处理液,培养细胞2-20天,小分子处理液是指:10%胎牛血清(Hyclone)+100U/ml青霉素(Sigma)+100μg/ml链霉素(Sigma)+高糖DMEM培养基(Gibco)+VPA(0.5mM~1.5mM)+Repsox(2~15uM),该培养***中10%胎牛血清也可以被血清替代品(invitrogen)以10%~20%的浓度替代;100U/ml青霉素(Sigma)和100μg/ml链霉素(Sigma)可以不使用。在37℃,5%CO2环境下培养细胞。
3、皮肤成纤维细胞的维持培养
上述第2步骤的处理结束后,完全更换为基础培养液,正常维持培养,在37℃,5%CO2环境下培养细胞。所述基础培养液:10%胎牛血清(Hyclone)+100U/ml青霉素(Sigma)+100μg/ml链霉素(Sigma)+高糖DMEM,中10%胎牛血清也可以被血清替代品(invitrogen)以10%~20%的浓度替代;100U/ml青霉素(Sigma)和100μg/ml链霉素(Sigma)可以不使用。
4、处理后细胞性状的鉴定
详细步骤同实施例1。
实施例5
第2步骤中皮肤成纤维细胞的小分子化合物处理为:更换基础培养液为小分子处理液,培养细胞2-20天,小分子处理液是指:10%胎牛血清(Hyclone)+100U/ml青霉素(Sigma)+100μg/ml链霉素(Sigma)+高糖DMEM培养基(Gibco)+VPA(0.5mM~1.5mM)+Repsox(2~15uM)+5-Aza-2′-deoxycytidine(0.5~20μM),该培养***中10%胎牛血清也可以被血清替代品(in vitrogen)以10%~20%的浓度替代;100U/ml青霉素(Sigma)和100μg/ml链霉素(Sigma)可以不使用。在37℃,5%CO2环境下培养细胞。其余步骤同实施例1。
实施例6
第2步骤中皮肤成纤维细胞的小分子化合物处理为:更换基础培养液为小分子处理液,培养细胞2-20天,小分子处理液是指:10%胎牛血清(Hyclone)+100U/ml青霉素(Sigma)+100μg/ml链霉素(Sigma)+高糖DMEM培养基(Gibco)+VPA(0.5mM~3mM),该培养***中10%胎牛血清也可以被血清替代品(invitrogen)以10%~20%的浓度替代;100U/ml青霉素(Sigma)和100μg/ml链霉素(Sigma)可以不使用。在37℃,5%CO2环境下培养细胞。其余步骤同实施例1。
实施例7
第2步骤中皮肤成纤维细胞的小分子化合物处理为:更换基础培养液为小分子处理液,培养细胞2-20天,小分子处理液是指:10%胎牛血清(Hyclone)+100U/ml青霉素(Sigma)+100μg/ml链霉素(Sigma)+高糖DMEM培养基(Gibco)+5-Aza-2′-deoxycytidine(0.2~20μM),该培养***中10%胎牛血清也可以被血清替代品(invitrogen)以10%~20%的浓度替代;100U/ml青霉素(Sigma)和100μg/ml链霉素(Sigma)可以不使用。在37℃,5%CO2环境下培养细胞。其余步骤同实施例1。
实施例8
第2步骤中皮肤成纤维细胞的小分子化合物处理为:更换基础培养液为小分子处理液,培养细胞2-20天,小分子处理液是指:10%胎牛血清(Hyclone)+100U/ml青霉素(Sigma)+100μg/ml链霉素(Sigma)+高糖DMEM培养基(Gibco)+RG108(0.5~30μM),该培养***中10%胎牛血清也可以被血清替代品(invitrogen)以10%~20%的浓度替代;100U/ml青霉素(Sigma)和100μg/ml链霉素(Sigma)可以不使用。在37℃,5%CO2环境下培养细胞。其余步骤同实施例1。
实施例9
第2步骤中皮肤成纤维细胞的小分子化合物处理为:更换基础培养液为小分子处理液,培养细胞2-20天,小分子处理液是指:10%胎牛血清(Hyclone)+100U/ml青霉素(Sigma)+100μg/ml链霉素(Sigma)+高糖DMEM培养基(Gibco)+Repsox(2~15uM)+CHIR99021(1μM~10μM)+VPA(0.5mM~1.5mM)+TTNPB(3μM~8μM)+AM580(0.03~0.08μM)+EPZ004777(3~8μM)+Y-27632(3~15μM)+L-Ascorbin acid 2-phosphate(0.15~0.25m M),该培养***中10%胎牛血清也可以被血清替代品(invitrogen)以10%~20%的浓度替代;100U/ml青霉素(Sigma)和100μg/ml链霉素(Sigma)可以不使用。在37℃,5%CO2环境下培养细胞。其余步骤同实施例1。
实施例10
第2步骤中皮肤成纤维细胞的小分子化合物处理为:更换基础培养液为小分子处理液,培养细胞2-20天,小分子处理液是指:10%胎牛血清(Hyclone)+100U/ml青霉素(Sigma)+100μg/ml链霉素(Sigma)+高糖DMEM培养基(Gibco)+forskolin(2μM~25μM)+Repsox(2~15uM)+CHIR99021(1μM~10μM)+VPA(0.5mM~1.5mM)+PDGF-AB(100~250ng/mL),该培养***中10%胎牛血清也可以被血清替代品(invitrogen)以10%~20%的浓度替代;100U/ml青霉素(Sigma)和100μg/ml链霉素(Sigma)可以不使用。在37℃,5%CO2环境下培养细胞。
其余步骤同实施例1。
实施例11
第2步骤中骨髓间充质干细胞的小分子化合物处理为:更换基础培养液为小分子处理液,培养细胞2-20天,小分子处理液是指:10%胎牛血清(Hyclone)+100U/ml青霉素(Sigma)+100μg/ml链霉素(Sigma)+高糖DMEM培养基(Gibco)+forskolin(2μM~25μM)+5-Aza-2′-deoxycytidine(0.5~20μM),该培养***中10%胎牛血清也可以被血清替代品(invitrogen)以10%~20%的浓度替代;100U/ml青霉素(Sigma)和100μg/ml链霉素(Sigma)可以不使用。在37℃,5%CO2环境下培养细胞。
其余步骤同实施例2。
上述实施例中细胞的变化见图1~图11;图1为小分子化合物作用后人源皮肤成纤维细胞的形态变化,A图未处理时,正常培养皮肤成纤维细胞,传到到第13代,一些原来呈长梭形生长的成纤维细胞,开始出现细胞体积变大,细胞形状不规则的老化情况,使用β-半乳糖苷酶染色,B图中可见衰老细胞染色成蓝色;使用如实施例1中的方法处理后,第14代的细胞,形态基本规则地呈长梭形,使用β-半乳糖苷酶染色,结果为阴性。
图2为小分子化合物作用后人源骨髓间充质干细胞形态的变化,A图未处理时,正常培养间充质干细胞,传到到第10代,一些原来呈梭形生长的细胞胞体变大,细胞拉长或者不规则型生长,使用β-半乳糖苷酶染色,B图中可见衰老细胞染色成蓝色;使用如实施例2中的方法处理后,第12代的细胞,形态基本规则地呈梭形,使用β-半乳糖苷酶染色,结果为阴性。
图3为小分子化合物作用后人源皮肤成纤维细胞细胞性状的鉴定,使用实施例1中的方法处理后,细胞和处理前一样,正常表达皮肤成纤维细胞的分子标志物CD29(A图),vimentin(B图);C图对比了处理前后细胞的增值情况,处理组Fib-treated的增值能力比未处理的Fib要明显。
图4为小分子化合物作用后人源骨髓间充质干细胞细胞性状的鉴定,如实施例2中,A图为处理前后间充质干细胞的分子标记物的表达,表达阳性的CD29,CD90,CD73以及阴性的CD34,CD45在处理前后的细胞中表达一致;B图中处理后的细胞MSC-treated要比处理前MSC的增值能力好;C图为处理后的MSC细胞,同样正常地进行3系分化,如图中显示了该细胞的成骨和成脂分化。
图5为通过小分子化合物作用后人源细胞的端粒长度的变化,A图中,以处理前的人源皮肤成纤维细胞作为对照,采用实施例1、实施例3~10中的方法,端粒长度增加且大于1.5倍;B图中处理了人源骨髓间充质干细胞,采用实施例2和实施例11的方法,处理后端粒长度增加。
图6为通过小分子化合物作用后人源细胞的端粒长度和年龄的对比,以及端粒相关基因的表达:与约35岁健康人群的平均端粒长度相比较,以及与自体相比较,52岁个体来源的皮肤成纤维细胞经过小分子组合处理后,其端粒长度增加明显,如A图所示;与端粒延长相关的基因TPP1瞬时高表达,如图B;图C显示,TERT(端粒酶反转录酶)在永生化细胞293T(“1”),Day12的Fib(人源皮肤成纤维细胞;“2”,实施例1),Day12的MSC(人源骨髓间充质干细胞;“3”,实施例2)中短暂表达。
图7为对小分子化合物作用后人源皮肤成纤维细胞中与衰老相关基因的表达变化,在处理后的细胞中,与衰老相关的基因均下调,例如与P53肿瘤抑制通路相关的基因CDKN1A、ATF3、GADD45B,BTG2,与衰老相关的H2A组蛋白家族成员X;
图8为小分子化合物作用后人源皮肤成纤维细胞体外功能活力的检测:如实施例3,皮肤成纤维细胞的I型胶原蛋白,在处理后明显上调;发育早期皮肤细胞的标记物ITGA6,在处理后上调明显。
图9为小分子化合物作用后人源皮肤成纤维细胞体内功能活力的检测:从皮肤分离的成纤维细胞具有较弱的免疫调节能力,通过实施例6的方法处理细胞后,在异体皮肤移植的模型中,将细胞培养48小时后的培养液注射到受体鼠体内,8天后观察皮肤移植部位,与对照组(未处理细胞组Fib,PBS组)相比,供体移植皮肤收到的免疫排斥反应较小,处理后的细胞具有明显的全身性免疫调节作用。
图10为对小分子化合物作用后长期传代的细胞干性的检测和成瘤性检测结果图:对实施例1中处理的细胞,进行Nod-SCID小鼠的皮下细胞移植,28天后未见成瘤,如图A白圈内所示;图B中对该组细胞进行干性基因OCT4的染色,未见表达。
图11小分子化合物作用后(如实施例1)人源皮肤成纤维细胞的克隆形成数量高于处理前,如图A所示;图B显示形成的克隆形态,处理前细胞(Fib)的克隆较小,细胞分散;处理过的细胞(Fib-treated)形态紧密,克隆较大。
Claims (2)
1.一种小分子化合物延长细胞端粒长度的体外培养方法,其特征在于,具体步骤如下:
(1)分离细胞,分离的细胞培养于基础培养液里;
(2)细胞传代大量扩增,待细胞处理的前一天,接种细胞密度1~2.5×104 /cm2培养于37℃,5%CO2的培养箱中;
(3)更换基础培养液为小分子处理液,在37℃,5%CO2环境下培养细胞2~20天,所述小分子处理液组成包括:10%胎牛血清+DMEM培养基+所述小分子化合物;
(4)更换小分子处理液为基础培养液,正常维持培养,在37℃,5%CO2环境下培养细胞;
所述小分子化合物选自下列任一项:
Forskolin+Repsox+CHIR99021+VPA+5-Aza-2 '-deoxycytidine;
Forskolin+Repsox+CHIR99021+VPA;
Forskolin+5-Aza-2 '-deoxycytidine;
VPA+Repsox;
VPA+Repsox+5-Aza-2 '-deoxycytidine;
VPA;
5-Aza-2 '-deoxycytidine;
RG108;
Repsox+CHIR99021+VPA+TTNPB+AM580+EPZ004777+Y-27632+L-Ascorbin acid 2-phosphate;
Forskolin+Repsox+CHIR99021+VPA+PDGF-AB;
Forskolin+5-Aza-2 '-deoxycytidine。
2.如权利要求1所述的方法,其特征在于,所述细胞来源于人的皮肤成纤维细胞或骨髓间充质干细胞。
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CN105579579A (zh) * | 2013-07-27 | 2016-05-11 | 高丽大学校产学协力团 | 包含小分子化合物的用于维持多能干细胞染色体稳定性的组合物 |
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-
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Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN105579579A (zh) * | 2013-07-27 | 2016-05-11 | 高丽大学校产学协力团 | 包含小分子化合物的用于维持多能干细胞染色体稳定性的组合物 |
CN104762324A (zh) * | 2015-03-06 | 2015-07-08 | 昆明学院 | 利用Runx2和小分子化合物诱导人成纤维细胞重编程为成骨细胞的方法 |
Non-Patent Citations (5)
Title |
---|
A XEN-like state bridges somatic cells to pluripotency during chemical reprogramming;Yang Zhao等;《Cell》;20151210;第163卷(第7期);摘要,第1678页右栏第一段,第1680页右栏最后1段-1第1683页左栏第1段,第1687页右栏第三段,第1689页右栏第三段,补充材料部分 * |
Molecules that promote or enhance reprogramming of somatic cells to induced pluripotent stem cells;Bo Feng 等;《Cell Stem Cell》;20090403;第301-312页 * |
Powering reprogramming with vitamin C;Yan Shi等;《Cell Stem Cell》;20100108;第6卷(第1期);第1-2页 * |
Yang Zhao等.A XEN-like state bridges somatic cells to pluripotency during chemical reprogramming.《Cell》.2015,第163卷(第7期), * |
提高细胞重编程效率和安全性的策略;虞欣璐等;《中国细胞生物学学报》;20161104;第38卷(第11期);第1394页右栏最后两段和1395页左栏第一段 * |
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