AbMole小課堂丨重組EGF在細胞培養(yǎng)、干細胞和類器官研究、細胞增殖和侵襲實驗中的應用表皮生長因子（EpidermalGrowthFactor,EGF）作為細胞生長因子家族的核心成員，通過與表皮生長因子受體（EGFR）結合激活下游信號通路，在細胞增殖、分化、存活及組織穩(wěn)態(tài)調控中發(fā)揮關鍵作用。EGF（重組EGF蛋白，AbMole，M9415）在多種細胞和動物模型中被廣泛應用，涉及基礎細胞生物學實驗、干細胞和類器官的培養(yǎng)等。EGF（RecombinantEGFProtein）的機理EGF（重組EGF蛋白，AbMole，M9415）是由53個氨基酸殘基組成的單鏈多肽，分子內(nèi)通過3對二硫鍵形成穩(wěn)定的空間結構，其N端區(qū)域是其與EGFR結合的關鍵結構域。在生理條件下，EGF主要由上皮細胞、成纖維細胞及免疫細胞分泌，以自分泌或旁分泌方式作用于靶細胞；EGF生物學效應的產(chǎn)生依賴于與細胞膜表面的EGFR（受體酪氨酸激酶家族成員之一）特異性結合，結合后EGFR發(fā)生二聚化，激活胞內(nèi)酪氨酸激酶結構域，引發(fā)自身酪氨酸殘基磷酸化；磷酸化的EGFR進一步招募下游適配分子（如Grb2、Shc），激活Ras/Raf/MEK/ERK、PI3K/Akt/mTOR及JAK/STAT等信號通路。其中，Ras/ERK通路主要調控細胞增殖與分化，PI3K/Akt通路參與細胞存活與代謝，JAK/STAT通路則與炎癥反應及細胞遷移相關。在實驗體系中，EGF可通過基因工程技術高效重組表達。重組EGF蛋白（RecombinantEGFProtein，Human，AbMole，M9415）因純度高、活性穩(wěn)定，已成為實驗研究用的EGF主要來源。圖SEQ圖\*ARABIC1.EGF家族成員及其各自受體ADDINEN.CITE<EndNote><Cite><Author>Abud</Author><Year>2021</Year><RecNum>977</RecNum><DisplayText><styleface="superscript">[1]</style></DisplayText><record><rec-number>977</rec-number><foreign-keys><keyapp="EN"db-id="f2td9w00a22awteprfrp9vaup9d9zwa9tdfr"timestamp="1756715250">977</key></foreign-keys><ref-typename="JournalArticle">17</ref-type><contributors><authors><author>Abud,HelenE</author><author>Chan,WingHei</author><author>Jardé,Thierry%JFrontiersincell</author><author>developmentalbiology</author></authors></contributors><titles><title>SourceandimpactoftheEGFfamilyofligandsonintestinalstemcells</title></titles><pages>685665</pages><volume>9</volume><dates><year>2021</year></dates><isbn>2296-634X</isbn><urls></urls></record></Cite></EndNote>[1]EGF（RecombinantEGFProtein）的研究應用EGF（RecombinantEGFProtein）用于細胞培養(yǎng)EGF（重組EGF蛋白，AbMole，M9415）是一些細胞（特別是原代細胞）培養(yǎng)基中重要的添加組分之一，例如在如小鼠皮膚角質形成細胞和支氣管上皮細胞的培養(yǎng)中，添加10-50ng/mL的EGF可顯著提高細胞貼壁率，促進細胞從G1期向S期過渡，延長細胞傳代壽命；在乳腺上皮細胞（MCF-10A）培養(yǎng)體系中，EGF與胰島素、Hydrocortisone協(xié)同作用，可維持細胞的正常上皮表型，抑制細胞衰老ADDINEN.CITE<EndNote><Cite><Author>Yusuf</Author><Year>2010</Year><RecNum>966</RecNum><DisplayText><styleface="superscript">[2]</style></DisplayText><record><rec-number>966</rec-number><foreign-keys><keyapp="EN"db-id="f2td9w00a22awteprfrp9vaup9d9zwa9tdfr"timestamp="1756708763">966</key></foreign-keys><ref-typename="JournalArticle">17</ref-type><contributors><authors><author>Yusuf,Rita</author><author>Frenkel,Krystyna%JCancerCellInternational</author></authors></contributors><titles><title>MorphologictransformationofhumanbreastepithelialcellsMCF-10A:dependenceonanoxidativemicroenvironmentandestrogen/epidermalgrowthfactorreceptors</title></titles><pages>30</pages><volume>10</volume><number>1</number><dates><year>2010</year></dates><isbn>1475-2867</isbn><urls></urls></record></Cite></EndNote>[2]。EGF（重組EGF蛋白）用于細胞遷移與侵襲實驗EGF（RecombinantEGFProtein，AbMole，M9415）還是重要的細胞趨化因子，可通過激活下游信號通路調控細胞骨架重排，促進細胞遷移與侵襲。EGF是Transwell遷移/侵襲實驗常用到一種生物活性分子?？稍赥ranswell小室上接種細胞（如MDA-MB-231乳腺癌細胞、HUVEC血管內(nèi)皮細胞），然后下室添加EGF（5-20ng/mL）作為趨化因子，培養(yǎng)24-48h后通過結晶紫染色或熒光標記計數(shù)遷移/侵襲到下室的細胞數(shù)量，可直觀評估EGF對細胞運動能力的促進作用ADDINEN.CITE<EndNote><Cite><Author>Biswenger</Author><Year>2018</Year><RecNum>968</RecNum><DisplayText><styleface="superscript">[3]</style></DisplayText><record><rec-number>968</rec-number><foreign-keys><keyapp="EN"db-id="f2td9w00a22awteprfrp9vaup9d9zwa9tdfr"timestamp="1756709372">968</key></foreign-keys><ref-typename="JournalArticle">17</ref-type><contributors><authors><author>Biswenger,Verena</author><author>Baumann,Nina</author><author>Jürschick,Johannes</author><author>H?ckl,Martina</author><author>Battle,Christopher</author><author>Schwarz,Jan</author><author>Horn,Elias</author><author>Zantl,Roman%JPloSone</author></authors></contributors><titles><title>CharacterizationofEGF-guidedMDA-MB-231cellchemotaxisinvitrousingaphysiologicalandhighlysensitiveassaysystem</title></titles><pages>e0203040</pages><volume>13</volume><number>9</number><dates><year>2018</year></dates><isbn>1932-6203</isbn><urls></urls></record></Cite></EndNote>[3]。EGF（RecombinantEGFProtein）用于干細胞和類器官培養(yǎng)EGF（RecombinantEGFProtein，AbMole，M9415）在干細胞培養(yǎng)中被廣泛用于促進細胞增殖和維持干細胞的干性。研究表明，EGF能夠激活下游的Ras/Raf/MAPK和PI3K/Akt等信號通路，從而促進干細胞的增殖和存活。例如，有研究發(fā)現(xiàn)在特定培養(yǎng)條件下，EGF與白血病抑制因子（LIF）協(xié)同作用，可顯著增強干細胞的增殖和多能性維持ADDINEN.CITEADDINEN.CITE.DATA[4]。表皮生長因子（EGF）還可誘導細胞的分化，例如EGF可刺激間充質干細胞（MSC）分化為骨形成細胞ADDINEN.CITE<EndNote><Cite><Author>Kratchmarova</Author><Year>2005</Year><RecNum>969</RecNum><DisplayText><styleface="superscript">[5]</style></DisplayText><record><rec-number>969</rec-number><foreign-keys><keyapp="EN"db-id="f2td9w00a22awteprfrp9vaup9d9zwa9tdfr"timestamp="1756710050">969</key></foreign-keys><ref-typename="JournalArticle">17</ref-type><contributors><authors><author>Kratchmarova,Irina</author><author>Blagoev,Blagoy</author><author>Haack-Sorensen,Mandana</author><author>Kassem,Moustapha</author><author>Mann,Matthias%JScience</author></authors></contributors><titles><title>Mechanismofdivergentgrowthfactoreffectsinmesenchymalstemcelldifferentiation</title></titles><pages>1472-1477</pages><volume>308</volume><number>5727</number><dates><year>2005</year></dates><isbn>0036-8075</isbn><urls></urls></record></Cite></EndNote>[5]。EGF還可以用于視網(wǎng)膜干細胞、神經(jīng)干細胞、腸道干細胞的分化誘導ADDINEN.CITE<EndNote><Cite><Author>Wang</Author><Year>2020</Year><RecNum>970</RecNum><DisplayText><styleface="superscript">[6]</style></DisplayText><record><rec-number>970</rec-number><foreign-keys><keyapp="EN"db-id="f2td9w00a22awteprfrp9vaup9d9zwa9tdfr"timestamp="1756710290">970</key></foreign-keys><ref-typename="JournalArticle">17</ref-type><contributors><authors><author>Wang,LX</author><author>Zhu,F</author><author>Li,JZ</author><author>Li,YL</author><author>Ding,XQ</author><author>Yin,J</author><author>Xiong,X</author><author>Yang,HS%JAnimal</author></authors></contributors><titles><title>EpidermalgrowthfactorpromotesintestinalsecretorycelldifferentiationinweaningpigletsviaWnt/β-cateninsignalling</title></titles><pages>790-798</pages><volume>14</volume><number>4</number><dates><year>2020</year></dates><isbn>1751-7311</isbn><urls></urls></record></Cite></EndNote>[6]。EGF（RecombinantEGFProtein，AbMole，M9415）在類器官培養(yǎng)中同樣發(fā)揮著重要作用，尤其是在維持干細胞干性和促進類器官形成方面。在多種類器官培養(yǎng)體系中，EGF被添加到培養(yǎng)基中，以支持細胞的增殖和分化。腸道類器官：在2009年，科學家Sato等成功建立了小鼠腸道類器官培養(yǎng)體系，該體系中添加了EGF、Noggin和R-spondin等因子。研究表明，這些因子能夠維持腸道干細胞的自我更新和分化，形成類似小腸的隱窩-絨毛樣復合體。此后，EGF被廣泛用于多種類器官的培養(yǎng)ADDINEN.CITE<EndNote><Cite><Author>Sato</Author><Year>2011</Year><RecNum>972</RecNum><DisplayText><styleface="superscript">[7]</style></DisplayText><record><rec-number>972</rec-number><foreign-keys><keyapp="EN"db-id="f2td9w00a22awteprfrp9vaup9d9zwa9tdfr"timestamp="1756710746">972</key></foreign-keys><ref-typename="JournalArticle">17</ref-type><contributors><authors><author>Sato,Toshiro</author><author>Stange,DanielE.</author><author>Ferrante,Marc</author><author>Vries,RobertG.J.</author><author>vanEs,JohanH.</author><author>vandenBrink,Stieneke</author><author>vanHoudt,WinanJ.</author><author>Pronk,Apollo</author><author>vanGorp,Joost</author><author>Siersema,PeterD.</author><author>Clevers,Hans</author></authors></contributors><titles><title>Long-termExpansionofEpithelialOrganoidsFromHumanColon,Adenoma,Adenocarcinoma,andBarrett'sEpithelium</title><secondary-title>Gastroenterology</secondary-title></titles><periodical><full-title>Gastroenterology</full-title><abbr-1>Gastroenterology</abbr-1></periodical><pages>1762-1772</pages><volume>141</volume><number>5</number><dates><year>2011</year></dates><publisher>Elsevier</publisher><isbn>0016-5085</isbn><urls><related-urls><url>/10.1053/j.gastro.2011.07.050</url></related-urls></urls><electronic-resource-num>10.1053/j.gastro.2011.07.050</electronic-resource-num><access-date>2025/09/01</access-date></record></Cite></EndNote>[7]。圖SEQ圖\*ARABIC2.腸道類器官的細胞組成ADDINEN.CITE<EndNote><Cite><Author>Sato</Author><Year>2011</Year><RecNum>972</RecNum><DisplayText><styleface="superscript">[7]</style></DisplayText><record><rec-number>972</rec-number><foreign-keys><keyapp="EN"db-id="f2td9w00a22awteprfrp9vaup9d9zwa9tdfr"timestamp="1756710746">972</key></foreign-keys><ref-typename="JournalArticle">17</ref-type><contributors><authors><author>Sato,Toshiro</author><author>Stange,DanielE.</author><author>Ferrante,Marc</author><author>Vries,RobertG.J.</author><author>vanEs,JohanH.</author><author>vandenBrink,Stieneke</author><author>vanHoudt,WinanJ.</author><author>Pronk,Apollo</author><author>vanGorp,Joost</author><author>Siersema,PeterD.</author><author>Clevers,Hans</author></authors></contributors><titles><title>Long-termExpansionofEpithelialOrganoidsFromHumanColon,Adenoma,Adenocarcinoma,andBarrett'sEpithelium</title><secondary-title>Gastroenterology</secondary-title></titles><periodical><full-title>Gastroenterology</full-title><abbr-1>Gastroenterology</abbr-1></periodical><pages>1762-1772</pages><volume>141</volume><number>5</number><dates><year>2011</year></dates><publisher>Elsevier</publisher><isbn>0016-5085</isbn><urls><related-urls><url>/10.1053/j.gastro.2011.07.050</url></related-urls></urls><electronic-resource-num>10.1053/j.gastro.2011.07.050</electronic-resource-num><access-date>2025/09/01</access-date></record></Cite></EndNote>[7]胃類器官：在胃類器官的研究中，EGF也被證明是維持胃干細胞增殖和分化所必需的。例如在從成體干細胞培養(yǎng)形成胃類器官時，需在培養(yǎng)基中添加EGF（一般為50ng/ml）ADDINEN.CITE<EndNote><Cite><Author>McCracken</Author><Year>2014</Year><RecNum>973</RecNum><DisplayText><styleface="superscript">[8]</style></DisplayText><record><rec-number>973</rec-number><foreign-keys><keyapp="EN"db-id="f2td9w00a22awteprfrp9vaup9d9zwa9tdfr"timestamp="1756711278">973</key></foreign-keys><ref-typename="JournalArticle">17</ref-type><contributors><authors><author>McCracken,KyleW</author><author>Catá,EmilyM</author><author>Crawford,CalynM</author><author>Sinagoga,KatieL</author><author>Schumacher,Michael</author><author>Rockich,BrianaE</author><author>Tsai,Yu-Hwai</author><author>Mayhew,ChristopherN</author><author>Spence,JasonR</author><author>Zavros,Yana%JNature</author></authors></contributors><titles><title>Modellinghumandevelopmentanddiseaseinpluripotentstem-cell-derivedgastricorganoids</title></titles><pages>400-404</pages><volume>516</volume><number>7531</number><dates><year>2014</year></dates><isbn>0028-0836</isbn><urls></urls></record></Cite></EndNote>[8]。并且胃腺體在這種含有EGF等多種生長因子的培養(yǎng)基中可生長為包含胃4種細胞譜系的三維類器官。培養(yǎng)基中除了需要添加EGF之外，還需加入Wnt-3a、Noggin（重組Noggin蛋白）、R-Spondin（R-spondin1，RSPO1蛋白）、FGF10（KGF-2）等細胞因子。這些因子共同營造適宜的微環(huán)境，調控細胞的增殖、分化和存活，促進胃類器官的正常生長和發(fā)育ADDINEN.CITE<EndNote><Cite><Author>McCracken</Author><Year>2014</Year><RecNum>973</RecNum><DisplayText><styleface="superscript">[8]</style></DisplayText><record><rec-number>973</rec-number><foreign-keys><keyapp="EN"db-id="f2td9w00a22awteprfrp9vaup9d9zwa9tdfr"timestamp="1756711278">973</key></foreign-keys><ref-typename="JournalArticle">17</ref-type><contributors><authors><author>McCracken,KyleW</author><author>Catá,EmilyM</author><author>Crawford,CalynM</author><author>Sinagoga,KatieL</author><author>Schumacher,Michael</author><author>Rockich,BrianaE</author><author>Tsai,Yu-Hwai</author><author>Mayhew,ChristopherN</author><author>Spence,JasonR</author><author>Zavros,Yana%JNature</author></authors></contributors><titles><title>Modellinghumandevelopmentanddiseaseinpluripotentstem-cell-derivedgastricorganoids</title></titles><pages>400-404</pages><volume>516</volume><number>7531</number><dates><year>2014</year></dates><isbn>0028-0836</isbn><urls></urls></record></Cite></EndNote>[8]。腫瘤類器官：EGF（重組EGF蛋白，AbMole，M9415）通過促進細胞增殖、分化和存活，為腫瘤類器官的形成和維持提供了重要的支持。例如在膠質母細胞瘤（GBM）類器官的培養(yǎng)中，使用含有EGF的培養(yǎng)基能夠顯著提高GBM類器官的培養(yǎng)成功率ADDINEN.CITE<EndNote><Cite><Author>Verduin</Author><Year>2023</Year><RecNum>976</RecNum><DisplayText><styleface="superscript">[9]</style></DisplayText><record><rec-number>976</rec-number><foreign-keys><keyapp="EN"db-id="f2td9w00a22awteprfrp9vaup9d9zwa9tdfr"timestamp="1756714694">976</key></foreign-keys><ref-typename="JournalArticle">17</ref-type><contributors><authors><author>Verduin,Maikel</author><author>Hoosemans,Linde</author><author>Vanmechelen,Maxime</author><author>vanHeumen,Mike</author><author>Piepers,JolandaAF</author><author>Astuti,Galuh</author><author>Ackermans,Linda</author><author>Schijns,OlafEMG</author><author>Kampen,KimR</author><author>Tjan-Heijnen,VivianneCG%JNeuro-oncologyadvances</author></authors></contributors><titles><title>Patient-derivedglioblastomaorganoidsreflecttumorheterogeneityandtreatmentsensitivity</title></titles><pages>vdad152</pages><volume>5</volume><number>1</number><dates><year>2023</year></dates><isbn>2632-2498</isbn><urls></urls></record></Cite></EndNote>[9]。研究表明，EGF還能夠顯著提高結直腸癌類器官的形成率，并且EGF可與N-Acetylcystein（NAC，AbMole）、FGF2、Y-27632等細胞因子或調節(jié)劑協(xié)同促進結直腸癌類器官的形成，為相關研究和藥物篩選提供了理想的模型ADDINEN.CITE<EndNote><Cite><Author>Otte</Author><Year>2019</Year><RecNum>974</RecNum><DisplayText><styleface="superscript">[10]</style></DisplayText><record><rec-number>974</rec-number><foreign-keys><keyapp="EN"db-id="f2td9w00a22awteprfrp9vaup9d9zwa9tdfr"timestamp="1756712160">974</key></foreign-keys><ref-typename="JournalArticle">17</ref-type><contributors><authors><author>Otte,J?rg</author><author>Dizdar,Levent</author><author>Behrens,Bianca</author><author>Goering,Wolfgang</author><author>Knoefel,WolframT</author><author>Wruck,Wasco</author><author>Stoecklein,NikolasH</author><author>Adjaye,James%JScientificReports</author></authors></contributors><titles><title>FGFsignallingintheself-renewalofcoloncancerorganoids</title></titles><pages>17365</pages><volume>9</volume><number>1</number><dates><year>2019</year></dates><isbn>2045-2322</isbn><urls></urls></record></Cite></EndNote>[10]。除了上述幾種類器官外，EGF也可用于肺類器官、甲狀腺類器官、肝臟類器官、唾液腺類器官、乳腺癌類器官，頭頸癌類器官的培養(yǎng)。三、范例詳解Heliyon.2024Aug27;10(17):e37079.武漢市第九醫(yī)院的科研人員在上述論文中探究了牙髓干細胞（DPSCs）的神經(jīng)分化，以及炎癥微環(huán)境中的分子機制，特別是ARMCX3基因在其中的作用。研究發(fā)現(xiàn)，ARMCX3在炎癥微環(huán)境中對DPSCs的神經(jīng)分化和炎癥反應有顯著影響，其機制可能涉及活性氧（ROS）信號通路。AbMole的bFGF（RecombinantHumanbFGFProtein，AbMole，M9406）和EGF（RecombinantHumanEGFProtein，AbMole，M9415）被用于支持DPSCs的神經(jīng)分化。具體地，bFGF和EGF被添加到培養(yǎng)基中，以促進DPSCs向神經(jīng)細胞分化。這些生長因子在培養(yǎng)基中的作用是提供必要的信號，以維持細胞的增殖和分化能力ADDINEN.CITE<EndNote><Cite><Author>Zhou</Author><Year>2024</Year><RecNum>490</RecNum><DisplayText><styleface="superscript">[11]</style></DisplayText><record><rec-number>490</rec-number><foreign-keys><keyapp="EN"db-id="f2td9w00a22awteprfrp9vaup9d9zwa9tdfr"timestamp="1752224437">490</key></foreign-keys><ref-typename="JournalArticle">17</ref-type><contributors><authors><author>Zhou,Quanying</author><author>Lei,Yi</author></authors></contributors><titles><title>ARMCX3regulatesROSsignaling,affectsneuraldifferentiationandinflammatorymicroenvironmentindentalpulpstemcells</title><secondary-title>Heliyon</secondary-title></titles><periodical><full-title>Heliyon</full-title><abbr-1>Heliyon</abbr-1></periodical><volume>10</volume><number>17</number><dates><year>2024</year></dates><publisher>Elsevier</publisher><isbn>2405-8440</isbn><urls><related-urls><url>/10.1016/j.heliyon.2024.e37079</url></related-urls></urls><electronic-resource-num>10.1016/j.heliyon.2024.e37079</electronic-resource-num><access-date>2025/07/11</access-date></record></Cite></EndNote>[11]。圖SEQ圖\*ARABIC3.ARMCX3knockdownfacilitatesneuraldifferentiationofhDPSCsADDINEN.CITE<EndNote><Cite><Author>Zhou</Author><Year>2024</Year><RecNum>490</RecNum><DisplayText><styleface="superscript">[11]</style></DisplayText><record><rec-number>490</rec-number><foreign-keys><keyapp="EN"db-id="f2td9w00a22awteprfrp9vaup9d9zwa9tdfr"timestamp="1752224437">490</key></foreign-keys><ref-typename="JournalArticle">17</ref-type><contributors><authors><author>Zhou,Quanying</author><author>Lei,Yi</author></authors></contributors><titles><title>ARMCX3regulatesROSsignaling,affectsneuraldifferentiationandinflammatorymicroenvironmentindentalpulpstemcells</title><secondary-title>Heliyon</secondary-title></titles><periodical><full-title>Heliyon</full-title><abbr-1>Heliyon</abbr-1></periodical><volume>10</volume><number>17</number><dates><year>2024</year></dates><publisher>Elsevier</publisher><isbn>2405-8440</isbn><urls><related-urls><url>/10.1016/j.heliyon.2024.e37079</url></related-urls></urls><electronic-resource-num>10.1016/j.heliyon.2024.e37079</electronic-resource-num><access-date>2025/07/11</access-date></record></Cite></EndNote>[11].參考文獻及鳴謝ADDINEN.REFLIST[1]HelenEAbud,WingHeiChan,Thierry%JFrontiersincellJardé,etal.,SourceandimpactoftheEGFfamilyofligandsonintestinalstemcells,9(2021)685665.[2]RitaYusuf,Kryst