Chinese Journal of Tissue Engineering Research ›› 2014, Vol. 18 ›› Issue (23): 3751-3755.doi: 10.3969/j.issn.2095-4344.2014.23.024
Previous Articles Next Articles
Huang Xia1,2, Pan Xing-hua1, Pang Rong-qing1, Ruan Guang-ping1, Cai Xue-min1
Revised:
2014-04-19
Online:
2014-06-04
Published:
2014-06-04
Contact:
Pan Xing-hua, M.D., Chief physician, Research Center for Stem Cells and Organ Tissue Engineering, Kunming General Hospital of Chengdu Military Area Command of Chinese PLA, Kunming 650032, Yunnan Province, China
About author:
Huang Xia, Research Center for Stem Cells and Organ Tissue Engineering, Kunming General Hospital of Chengdu Military Area Command of Chinese PLA, Kunming 650032, Yunnan Province, China; Clinical School, Kunming Medical University, Kunming 650031, Yunnan Province, China
Supported by:
he National Natural Science Foundation of China, No. 31172170
CLC Number:
Huang Xia, Pan Xing-hua, Pang Rong-qing, Ruan Guang-ping, Cai Xue-min. Umbilical cord-derived mesenchymal stem cell culture: dyeing and tracer technique[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(23): 3751-3755.
2.1 纳入文献基本情况 初检得到203篇文献,其中英文文献193篇,中文文献10篇。阅读标题和摘要进行初筛,排除与研究目的不符和重复性文章;查阅全文,判断与纳入标准一致的文章,最后选择35篇符合标准的文献。文献[1-11]研究了脐带间充质干细胞基本理论知识,文献[12-23]研究了脐带间充质干细胞的培养及培养条件的优化,文献[24-35]篇研究了脐带间充质干细胞的体外荧光染料标记。 2.2 结果描述 2.2.1 脐带间充质干细胞的培养 脐带标本应选择健康足月产出生的新生儿脐带。分离培养脐带间充质干细胞主要有酶消化法[12]、组织平铺法等[13]。①酶消化法:将脐带组织剪碎,用Ⅳ型胶原酶消化后离心,取下层沉淀,用胰蛋白酶消化后再次离心,取沉淀,以PBS吹打悬浮,100 μm滤网过滤,滤过液以2 000 r/min离心 10 min,取沉淀,以PBS洗涤细胞2遍,每遍800 r/min离心10 min,以1×109 L-1的细胞浓度悬浮于DMEM培养基中,置于培养箱培养。②组织平铺法[14]:分离培养细胞。将组织块剪成1 mm×1 mm×1 mm大小,平铺于培养瓶底,组织块接种至DMEM/F12培养基中,放置于二氧化碳培养箱内培养,1周之后更换培养基。细胞生长至80%融合时,用0.25%胰酶将其消化,进行传代培养[14]。培养基是关系细胞培养成功与否的一个十分重要因素[15],目前用于培养脐带间充质干细胞的培养基大多为L-DMEM培养基加上体积分数10%胎牛血清[15]。 2.2.2 脐带间充质干细胞的培养条件优化 在从脐带上分离间充质干细胞过程中,酶处理在分离脐带间充质干细胞中起十分重要作用[16]。目前使用的主要有组织块贴壁法和酶消化法或两者的结合。使用组织块贴壁法分离时[17-18],7-10 d后可见贴壁生长的单个长条梭形细胞从组织块中分离出来,细胞融合可以达80%-90%时,用0.25%的胰蛋白酶进行消化传代[17-18]。酶消化法,就是对剪碎后的脐带组织块加入胰蛋白酶、胶原酶或各种酶的混合液对其进行消化,直接得到贴壁生长的单个细胞。在消化过程中加入DNA酶可有助于分散细胞[19]。虽然酶消化法能较快分离出间充质干细胞,但是酶体系中可能会降解细胞膜外层蛋白,甚至可能导致细胞较难贴壁。因此,把握好酶的用法、用量及消化的时间是分离脐带间充质干细胞的关键。 目前,已有多个研究小组已成功从脐带组织中分离出具有间充质干细胞特性的干细胞。据研究组和培养方法的差异,将所得到的贴壁细胞分成为间充质干细胞、类间充质干细胞或基质干细胞[20]。研究发现脐带血管周围的脐带沃顿胶组织中含有大量的间充质干细胞,通过植块法或酶消化法进行培养可以得到更高增殖活性的具有间充质干细胞性质的干细胞[21]。Kern等[21]认为,原因可能是培养体系的差别影响了细胞的增殖能力,但本体系中细胞平均4 d即可传1代,细胞平均倍增6.18倍,因此短期培养也可获得足够的细胞数进行细胞临床等方面的应用。利用流式细胞仪检测发现,将细胞传至3代之后,获得的间充质干细胞纯度与原有的纯度有所提高,即细胞表达间充质干细胞的特性[18,21-22]。 齐凯等[23]比较了以Ⅱ型胶原酶为主要成分的混合酶Ⅰ、以胰酶为主要成分的混合酶Ⅱ和以Ⅱ型胶原酶-胰酶-DNA酶为主要成分的混合酶Ⅲ对脐带间充质干细胞分离效果的影响,结果显示以Ⅱ型胶原酶-胰酶-DNA酶为主要成分的混合酶Ⅲ可以增高从脐静脉酶解得到的总细胞数、培养3代后得到的脐带间充质干细胞细胞数,虽然在酶降解3 h后得到的细胞中死细胞比较多,但是经体外培养传代后对其增殖能力没有影响。齐凯 等[23]认为混合酶Ⅲ对分离脐带间充质干细胞有利。 2.2.3 脐带间充质干细胞的体外荧光染料标记 荧光染料标记在医学、生命科学等方面具有相当大的应用前景和潜在价值。随着医学生命科学的向前发展,以及计算机技术、激光技术、荧光光谱测定技术的不断进步,很多染料特别是荧光染料在DNA测序、毒物分析、细胞检测、临床诊断等多方面得到了很好的应用。荧光染料标记的原理是将一些能够结合到胞内或胞膜不同组织上的荧光染料,从而达到对细胞进行观察的目的。在脐带间充质干细胞中常用的荧光染料主要有DAPI、Di-I、PKH26/PKH67、CFSE、绿色荧光蛋白等[24-27]。 DAPI:DAPI即4’,6-二脒基-2-苯基吲哚(4’,6-diamidino-2-phenylindole)[24,28],相对分子质量350。它具有专一性强、灵敏度高、稳定性好等特点。是一种能够与DNA强力结合的荧光染料,常利用荧光显微镜进行检测。由于DAPI能够透过完整的胞膜,它能使用于活细胞和固定细胞的染色标记。在紫外光的激发下发出浅蓝色荧光。但是随着细胞的不断分裂增殖,DAPI被均匀分配到子代细胞中,以至于DAPI标记强度降低,灵敏度随之下降。DAPI除与DNA双链相结合之外,还可以和胞浆之中的微管蛋白相结合,所以细胞浆也能标记成蓝色。DAPI标记细胞的缺点就是如果标记的细胞死亡后,就能释放出来DAPI,把周围没有标记的细胞标记,导致假阳性,上述问题是人们使用此类荧光染料标记细胞时该考虑的问题。Castanheira等[28]评估DAPI作为核示踪剂标记间充质干细胞,观察将其注射至视网膜损伤的大鼠视网膜玻璃体腔内的迁移和分布。Castanheira等认为DAPI标记干细胞在体内实验中不是一个理想的标记物。由于随着细胞的分裂增殖,DAPI 被平分至子代细胞中,引起DAPI标记强度下降,灵敏度减低。因此,DAPI也只适用于短期的标记示踪。 Di-I:Di-I (1,1’-dioctacecyl-3,3,3’,3’-tetramethylindoc-yanine perchlorate) 即1,1’-双十八烷-3,3,3’,3’ -四甲基吲哚碳花青-高氯酸盐,是一种亲脂性长链碳花青染料,易嵌入生物质膜内并在膜内作侧向扩散运动,从而标记整个细胞[25,29]。还可以通过活细胞的胞饮作用进入胞质,标记整个细胞质。在水中具有弱荧光,当掺入膜中时荧光增强,而且光稳定性十分好,荧光保持时间长,在标记细胞内消失慢,不容易在标记与非标记细胞之间传递,无细胞毒性,在细胞膜中的存在不影响细胞活力、发育或其他生理学性质。Hu等[29]用CM-DiI标记间充质干细胞,观察到CM-DiI标记可维持到移植后8周。Dil标记技术简单,染色快,标记效率也很高。但因为Di-I标记的是细胞膜,因此存在着随着细胞分裂荧光强度减低的缺点,不适于长期细胞的追踪。 PKH26/PKH67:PKH26/PKH67 PKH[26,30]荧光细胞标记试剂细胞膜标记技术,在细胞膜的脂质双分子层中稳定结合红色荧光染料-PKH26/绿色荧光染料- PKH67,产生稳定、清晰、精确和可重复的荧光标记细胞,可广泛用于动物、植物细胞和其它含颗粒胞膜的标记。在适当的标记条件下,PKH26不影响细胞的增殖和分化能力;当细胞分裂时,PKH26可以平分至两个子代细胞中,荧光强度是上一代细胞的一半,胞膜能发出红色荧光,不会影响胞膜表面标记物的表达,在恰当的条件下不影响细胞的活力。Shao-Fang等[26]探讨PKH26标记脐带间充质干细胞是否影响细胞的形态、表型、核扩散和分泌能力。分离出来的脐带间充质干细胞用PKH26标记,利用显微镜观察细胞的形态。没有检测到PKH26标记与未标记的细胞存在细胞形态学、细胞生长和增殖效率的差异。Shao-Fang等认为PKH26标记的荧光强度随着时间的推移逐渐降低。总之,PKH26标记人类脐带间充质干细胞是一种安全、有效的标记方式。因此,PKH经常被应用于体内、体外短期的细胞示踪研究实验。 羧基荧光素乙酰乙酸(CFSE):CFSE是一种能够穿过胞膜的荧光染料,进入细胞后不可逆地与胞内具有非酶促水解作用的梭基荧光素二醋酸盐基团结合,偶联到细胞蛋白质上,且不会引起细胞发生凋亡或死亡[27,31-32]。CFSE一旦进入细胞后就不能从细胞中释放出来,并且自发地不可逆地与胞内蛋白质和细胞膜表面蛋白结合。当细胞进行有丝分裂和细胞增殖时,具有荧光的胞质蛋白被平分到第二代细胞中,这样与第一代细胞相比,这样荧光强度也将会减弱至原来的一半;按这样计算,有丝分裂至第三代细胞的荧光强度将会比第二代细胞更加减弱。此现象可以在488 nm的激发光下,使用流式细胞仪进行检测分析,通过检测到细胞荧光强度不断的下降,详细分析得出细胞分裂和增殖的情况。所以,CFSE经常被用来做活细胞检测实验和用荧光电镜观察细胞长期活动的实验。Chadli等[31]研究了使用胞内荧光染料CFSE检测正常人类角质细胞的细胞分裂。结果表明,CFSE标记的细胞能跟踪监测的角化细胞响应性增长,在这里充分体现了转化生长因子β1介导的细胞循环抑制。因此,CFSE常被用来做活细胞检测试验和用荧光电镜观察细胞长期活动的试验。 绿色荧光蛋白:绿色荧光蛋白是一类存在于腔肠动物体内的生物发光蛋白,由于水母整体所发出的荧光及提取的蛋白质颗粒所发出的荧光都呈绿色,因此,把这种蛋白质命名为绿色荧光蛋白[33]。绿色荧光蛋白是能在异源细胞内表达后,能自发产生荧光的蛋白,且绿色荧光蛋白的相对分子质量比较小,N-端和C-端都能接受蛋白的融合,是很好的标记物,可以进行活细胞实时定位观察,更加能靠近自然真实的状态。如果在活细胞中直接观察蛋白质向细胞核、内质网运动的状态,并且可以实时观察到外界信号刺激下,目的蛋白质的变化过程,使用荧光显微镜观察,使研究更加方便。借助激光共聚焦显微镜,其图像效果更好,利用现代的计算机软件,可以进行三维图像的显示[33]。刘毅等[34]构建共表达增强型绿色荧光蛋白基因和人胰岛素(insulin)基因的慢病毒载体,探讨其对人脐带间充质干细胞的转染情况。结果发现成功构建了共表达insulin基因和增强型绿色荧光蛋白基因的重组慢病毒载体pLenti6.3-insulin- IRES-EGFP,并对其成功包装、纯化及浓缩,病毒滴度为1.3×108 TU/mL。不同MOI的重组慢病毒感染人脐带间充质干细胞之后,通过绿色荧光蛋白表达的阳性细胞数筛选其最适MOI为10,此时对细胞的转染率可以达到90%[34]。刘毅等[34]认为构建的携带insulin基因的重组慢病毒载体pLenti6.3-insulin-IRES-EGFP可有效转染人脐带间充质干细胞,表达insulin蛋白。唐莉等[35]将携带绿色荧光蛋白的慢病毒感染hU-CMSCs,并且观察其对Oct4表达的影响。结果发现体外培养出的人脐带间充质干细胞呈长梭形成纤维细胞样,荧光显微镜观察在感染96 h后荧光表达最强;当以MOI=20感染细胞96 h后,绿色荧光蛋白阳性率达75%以上;与未感染组相比,MTT显示绿色荧光蛋白慢病毒对细胞增殖没有明显影响(P > 0.05)。免疫荧光染色检查显示Oct4在培养的第2周、第8周中都有表达且定位于细胞核。唐莉等[35]认为慢病毒携带的绿色荧光蛋白基因能够表达于人脐带间充质干细胞中,并且不影响Oct4基因的表达。"
[1] Löhle M, Hermann A, Glaß H, et al. Differentiation efficiency of induced pluripotent stem cells depends on the number of reprogramming factors. Stem Cells. 2012;30(3):570-579. [2] Gifford Casey A, Ziller Michael J, Gu H, et al. Transcriptional and epigenetic dynamics during specification of human embryonic stem cells. Cell. 2013;153(5):1149-1163. [3] Sanchez-Adams J, Athanasiou KA. Dermis isolated adult stem cells for cartilage tissue engineering. Biomaterials. 2012; 33(1):109-119. [4] Jung Y, Bauer G, Nolta JA. Concise review: Induced pluripotent stem cell-derived mesenchymal stem cells: progress toward safe clinical products. Stem Cells. 2012; 30(1): 42-47. [5] Hao L, Sun HQ, Guo XS, et al. Exogenous gene expression in vitro and in vivo in bone marrow mesenchymal stem cells modified by hPDGF-A and hBD(2). Zhongguo Shi Yan Xue Ye Xue Za Zhi.2009;17(3):685-689. [6] Chen Y, Yu B, Xue G, et al. Effects of storage solutions on the viability of human umbilical cord mesenchymal stem cells for transplantation. Cell Transplant. 2013;22(6):1075-1086. [7] Romanov YA, Svintsitskaya VA, Smirnov VN. Searching for alternative sources of postnatal human mesenchymal stem cells: candidate MSC-like cells from umbilical cord. Stem Cells. 2003;21(1):105-110. [8] Kim DW, Staples M, Shinozuka K, et al. Wharton's Jelly-Derived Mesenchymal Stem Cells: Phenotypic Characterization and Optimizing Their Therapeutic Potential for Clinical Applications. Int J Mol Sci. 2013;14(6):11692- 11712. [9] Wang HS, Hung SC, Peng ST, et al. Mesenchymal stem cells in the Wharton's jelly of the human umbilical cord. Stem Cells. 2004;22(7):1330-1337. [10] Sarugaser R, Lickorish D, Baksh D, et al. Human umbilical cord perivascular (HUCPV) cells: a source of mesenchymal progenitors. Stem Cells. 2005;23(2):220-229. [11] Lu LL, Song YP, Wei XD, et al. Comparative characterization of mesenchymal stem cells from human umbilical cord tissue and bone marrow. Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2008;16(1):140-146. [12] Kestendjieva S, Kyurkchiev D, Tsvetkova G, et al. Characterization of mesenchymal stem cells isolated from the human umbilical cord. Cell Biol Int. 2008;32(7):724-732. [13] Ma L, Feng XY, Cui BL, et al. Human umbilical cord Wharton's Jelly-derived mesenchymal stem cells differentiation into nerve-like cells. Chin Med J (Engl). 2005;118(23):1987-1993. [14] 庞荣清,何洁,李福兵,等.一种简单的人脐带间充质干细胞分离培养方法[J].中华细胞与干细胞杂志(电子版), 2011,1(2):30-33. [15] Mannello F, Tonti GA. Concise review: no breakthroughs for human mesenchymal and embryonic stem cell culture: conditioned medium, feeder layer, or feeder-free; medium with fetal calf serum, human serum, or enriched plasma; serum-free, serum replacement nonconditioned medium, or ad hoc formula? All that glitters is not gold!. Stem Cells. 2007;25(7):1603-1609. [16] Tsagias N, Koliakos I, Karagiannis V, et al. Isolation of mesenchymal stem cells using the total length of umbilical cord for transplantation purposes. Transfus Med. 2011; 21(4):253-261. [17] Diao Y, Ma Q, Cui F, et al. Human umbilical cord mesenchymal stem cells: osteogenesis in vivo as seed cells for bone tissue engineering. Journal of biomedical materials research Part A. 2009;91(1):123-131. [18] Ishige I, Nagamura-Inoue T, Honda MJ, et al. Comparison of mesenchymal stem cells derived from arterial, venous, and Wharton's jelly explants of human umbilical cord. Int J Hematol. 2009; 90(2):261-269. [19] Schultz SS, Lucas PA. Human stem cells isolated from adult skeletal muscle differentiate into neural phenotypes. J Neurosci Methods. 2006;152(1-2):144-155. [20] 徐燕,李长虹,孟恒星,等.人脐带间充质干细胞分离培养条件的优化及其生物学特性[J].中国组织工程研究与临床康复,2009, 13(32): 6289-6294. [21] Kern S, Eichler H, Stoeve J, et al. Comparative analysis of mesenchymal stem cells from bone marrow, umbilical cord blood, or adipose tissue. Stem Cells. 2006;24(5):1294-1301. [22] Seshareddy K, Troyer D, Weiss ML. Method to isolate mesenchymal-like cells from Wharton's Jelly of umbilical cord. Methods Cell Biol. 2008;86:101-119. [23] 齐凯,董丽媛,陈显久,等.人脐带来源间充质干细胞分离培养方法的优化[J].中国组织工程研究与临床康复,2011,15(23): 4220-4224. [24] Chazotte B. Labeling nuclear DNA using DAPI. Cold Spring Harbor protocols. 2011;2011(1):pdb prot5556. [25] Hu KX, Wang MH, Fan C, et al. CM-DiI labeled mesenchymal stem cells homed to thymus inducing immune recovery of mice after haploidentical bone marrow transplantation. Int Immunopharmacol. 2011;11(9):1265-1270. [26] Shao-Fang Z, Hong-Tian Z, Zhi-Nian Z, et al. PKH26 as a fluorescent label for live human umbilical mesenchymal stem cells. In Vitro Cell Dev Biol Anim. 2011;47(8):516-520. [27] Godfrey WR, Krampf MR, Taylor PA, et al. Ex vivo depletion of alloreactive cells based on CFSE dye dilution, activation antigen selection, and dendritic cell stimulation. Blood. 2004; 103(3):1158-1165. [28] Castanheira P, Torquetti LT, Magalhas DR, et al. DAPI diffusion after intravitreal injection of mesenchymal stem cells in the injured retina of rats. Cell Transplant. 2009;18(4): 423-431. [29] Schmidt H, Rathjen FG. DiI-labeling of DRG neurons to study axonal branching in a whole mount preparation of mouse embryonic spinal cord. J Vis Exp. 2011;(58):pii: 3667. [30] Yang Q, Peng J, Guo Q, et al. A cartilage ECM-derived 3-D porous acellular matrix scaffold for in vivo cartilage tissue engineering with PKH26-labeled chondrogenic bone marrow-derived mesenchymal stem cells. Biomaterials. 2008;29(15):2378-2387. [31] Chadli L, Cadio E, Vaigot P, et al. Monitoring the cycling activity of cultured human keratinocytes using a CFSE-based dye tracking approach. Methods Mol Biol. 2013;989:83-97. [32] Urbani S, Caporale R, Lombardini L, et al. Use of CFDA-SE for evaluating the in vitro proliferation pattern of human mesenchymal stem cells. Cytotherapy. 2006;8(3):243-253. [33] Patterson GH, Lippincott-Schwartz J. A photoactivatable GFP for selective photolabeling of proteins and cells. Science. 2002;297(5588):1873-1877. [34] Liu Y, Xue M. Recombinant human insulin gene lentivirus transfecting human umbilical cord mesenchymal stem cells in vitro[J]. Zhongguo xiu fu chong jian wai ke za zhi. 2010;24(7): 822-827. [35] Tang L, Chang J. Effect of GFP-containing lentivirus infection on the expression of octamer transcription factor 4 in human umbilical cord mesenchymal stem cells. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2013;29(3):292-296. |
[1] | Lin Qingfan, Xie Yixin, Chen Wanqing, Ye Zhenzhong, Chen Youfang. Human placenta-derived mesenchymal stem cell conditioned medium can upregulate BeWo cell viability and zonula occludens expression under hypoxia [J]. Chinese Journal of Tissue Engineering Research, 2021, 25(在线): 4970-4975. |
[2] | Pu Rui, Chen Ziyang, Yuan Lingyan. Characteristics and effects of exosomes from different cell sources in cardioprotection [J]. Chinese Journal of Tissue Engineering Research, 2021, 25(在线): 1-. |
[3] | Liu Zhichao, Zhang Fan, Sun Qi, Kang Xiaole, Yuan Qiaomei, Liu Genzhe, Chen Jiang. Morphology and activity of human nucleus pulposus cells under different hydrostatic pressures [J]. Chinese Journal of Tissue Engineering Research, 2021, 25(8): 1172-1176. |
[4] | Zhang Xiumei, Zhai Yunkai, Zhao Jie, Zhao Meng. Research hotspots of organoid models in recent 10 years: a search in domestic and foreign databases [J]. Chinese Journal of Tissue Engineering Research, 2021, 25(8): 1249-1255. |
[5] | Wang Zhengdong, Huang Na, Chen Jingxian, Zheng Zuobing, Hu Xinyu, Li Mei, Su Xiao, Su Xuesen, Yan Nan. Inhibitory effects of sodium butyrate on microglial activation and expression of inflammatory factors induced by fluorosis [J]. Chinese Journal of Tissue Engineering Research, 2021, 25(7): 1075-1080. |
[6] | Wang Xianyao, Guan Yalin, Liu Zhongshan. Strategies for improving the therapeutic efficacy of mesenchymal stem cells in the treatment of nonhealing wounds [J]. Chinese Journal of Tissue Engineering Research, 2021, 25(7): 1081-1087. |
[7] | Liao Chengcheng, An Jiaxing, Tan Zhangxue, Wang Qian, Liu Jianguo. Therapeutic target and application prospects of oral squamous cell carcinoma stem cells [J]. Chinese Journal of Tissue Engineering Research, 2021, 25(7): 1096-1103. |
[8] | Xie Wenjia, Xia Tianjiao, Zhou Qingyun, Liu Yujia, Gu Xiaoping. Role of microglia-mediated neuronal injury in neurodegenerative diseases [J]. Chinese Journal of Tissue Engineering Research, 2021, 25(7): 1109-1115. |
[9] | Li Shanshan, Guo Xiaoxiao, You Ran, Yang Xiufen, Zhao Lu, Chen Xi, Wang Yanling. Photoreceptor cell replacement therapy for retinal degeneration diseases [J]. Chinese Journal of Tissue Engineering Research, 2021, 25(7): 1116-1121. |
[10] | Jiao Hui, Zhang Yining, Song Yuqing, Lin Yu, Wang Xiuli. Advances in research and application of breast cancer organoids [J]. Chinese Journal of Tissue Engineering Research, 2021, 25(7): 1122-1128. |
[11] | Wang Shiqi, Zhang Jinsheng. Effects of Chinese medicine on proliferation, differentiation and aging of bone marrow mesenchymal stem cells regulating ischemia-hypoxia microenvironment [J]. Chinese Journal of Tissue Engineering Research, 2021, 25(7): 1129-1134. |
[12] | Zeng Yanhua, Hao Yanlei. In vitro culture and purification of Schwann cells: a systematic review [J]. Chinese Journal of Tissue Engineering Research, 2021, 25(7): 1135-1141. |
[13] | Kong Desheng, He Jingjing, Feng Baofeng, Guo Ruiyun, Asiamah Ernest Amponsah, Lü Fei, Zhang Shuhan, Zhang Xiaolin, Ma Jun, Cui Huixian. Efficacy of mesenchymal stem cells in the spinal cord injury of large animal models: a meta-analysis [J]. Chinese Journal of Tissue Engineering Research, 2021, 25(7): 1142-1148. |
[14] | Hou Jingying, Yu Menglei, Guo Tianzhu, Long Huibao, Wu Hao. Hypoxia preconditioning promotes bone marrow mesenchymal stem cells survival and vascularization through the activation of HIF-1α/MALAT1/VEGFA pathway [J]. Chinese Journal of Tissue Engineering Research, 2021, 25(7): 985-990. |
[15] | Shi Yangyang, Qin Yingfei, Wu Fuling, He Xiao, Zhang Xuejing. Pretreatment of placental mesenchymal stem cells to prevent bronchiolitis in mice [J]. Chinese Journal of Tissue Engineering Research, 2021, 25(7): 991-995. |
Viewed | ||||||
Full text |
|
|||||
Abstract |
|
|||||