細(xì)胞處理(細(xì)胞制造2025系列3)

1、細(xì)胞處理（細(xì)胞制造2025系列3）口細(xì)胞處理細(xì)胞在體外人工環(huán)境下生長(zhǎng)，是制造過程中的一個(gè)明確步驟。細(xì)胞培養(yǎng)的每個(gè)參數(shù)，包括容器，培養(yǎng)基，營(yíng)養(yǎng)物和物理化學(xué)環(huán)境，都將影響治療有效性所需的細(xì)胞特性。新興細(xì)胞醫(yī)療產(chǎn)品的發(fā)展需要大規(guī)模高速度的細(xì)胞制造科技技術(shù)。然而，目前方法所需的物料，空間，勞動(dòng)力和時(shí)間都阻礙了現(xiàn)有工藝滿足這種日益增長(zhǎng)的需求。為了實(shí)現(xiàn)在未來年內(nèi)大規(guī)模生產(chǎn)高品質(zhì)的細(xì)胞，細(xì)胞制造產(chǎn)業(yè)必須共同開發(fā)，優(yōu)化和應(yīng)用更廉價(jià)有效的細(xì)胞處理科技。提高細(xì)胞生產(chǎn)過程的一致性和可靠性，同樣可以提高細(xì)胞產(chǎn)品的質(zhì)量和功效。當(dāng)前的挑戰(zhàn)為了實(shí)現(xiàn)大規(guī)模，低成本，可持續(xù)生產(chǎn)高質(zhì)量的細(xì)胞，細(xì)胞制造產(chǎn)業(yè)必須努力克服細(xì)胞處理過程

2、中遇到的以下挑戰(zhàn)：現(xiàn)有培養(yǎng)平臺(tái)的線性特征目前大多數(shù)培養(yǎng)平臺(tái)（例如，平面培養(yǎng)系統(tǒng)或飼養(yǎng)細(xì)胞）都是線性的并需要更大的表面積去大量的生產(chǎn)細(xì)胞。當(dāng)前培養(yǎng)系統(tǒng)所導(dǎo)致的空間限制，加上所需的大量培養(yǎng)基和勞動(dòng)力，使得現(xiàn)有的培養(yǎng)方法在產(chǎn)業(yè)化規(guī)模制造中，在經(jīng)濟(jì)上是不切實(shí)際的。與此同時(shí)，線性培養(yǎng)系統(tǒng)很難控制營(yíng)養(yǎng)梯度，難以實(shí)現(xiàn)一致性的細(xì)胞特征和質(zhì)量。開放式的培養(yǎng)環(huán)境增加了污染的可能性在常規(guī)研究環(huán)境中，目前分離細(xì)胞以便于更進(jìn)一步擴(kuò)增的操作是在開放式系統(tǒng)中進(jìn)行的，容器是暴露于周圍的環(huán)境之中的。每一次打開培養(yǎng)容器都將置于霉菌、酵母、病毒、支原體和其他細(xì)胞系污染的潛在風(fēng)險(xiǎn)之下。為保持無菌環(huán)境以減少這種風(fēng)險(xiǎn)而進(jìn)行的維護(hù)都是時(shí)間

3、和勞動(dòng)力密集型工作。研發(fā)封閉式培養(yǎng)系統(tǒng)技術(shù)，完全封閉與外部環(huán)境，是目前迫切的需求。（推薦嘗試英普樂孚生物技術(shù)（上海）有限公司研發(fā)的全封閉式密閉培養(yǎng)系統(tǒng)，包含：袋裝培養(yǎng)基、細(xì)胞培養(yǎng)袋、細(xì)胞轉(zhuǎn)移袋、細(xì)胞保存袋等。通過專利管路連接系統(tǒng)，實(shí)現(xiàn)流程全封閉式操作，避免開放式操作環(huán)境造成污染的可能性。）細(xì)胞難以理解的復(fù)雜性細(xì)胞制造產(chǎn)業(yè)對(duì)人類細(xì)胞，特別是干細(xì)胞和免疫細(xì)胞的生物學(xué)和應(yīng)激特性缺乏足夠了解。工業(yè)化生產(chǎn)細(xì)胞不同于細(xì)胞在體內(nèi)的狀態(tài)，使其更復(fù)雜更加難以被理解。由于細(xì)胞固有的復(fù)雜性，難以確定其所需的特性，或者操作模式，如何使細(xì)胞在治療上有用，制造具有這些特性的細(xì)胞所需的培養(yǎng)環(huán)境和分化過程。成本高、有限供應(yīng)和

4、材料的不一致性培養(yǎng)基包括生長(zhǎng)因子，營(yíng)養(yǎng)物質(zhì)和試劑通常是細(xì)胞制造過程中最昂貴的部分。高成本，及一些材料供應(yīng)及保存期限的限制，阻礙了現(xiàn)有培養(yǎng)平臺(tái)滿足不斷增長(zhǎng)的細(xì)胞生產(chǎn)需求。更復(fù)雜的情況是，一些培養(yǎng)基組份，特別是動(dòng)物血清，具有明顯的批間差異，將會(huì)降低所生產(chǎn)細(xì)胞特性的一致性，甚至污染細(xì)胞產(chǎn)品。細(xì)胞分離方法效率低目前許多細(xì)胞處理系統(tǒng)，細(xì)胞必須從培養(yǎng)容器的表面被分離并轉(zhuǎn)移到具有新鮮培養(yǎng)基的新容器中進(jìn)一步增殖擴(kuò)增。當(dāng)處理最終產(chǎn)品時(shí)，細(xì)胞必須與培養(yǎng)基，未分化的細(xì)胞和其他顆粒物分離，以獲得最終產(chǎn)品所需的細(xì)胞純度。當(dāng)前的分離方法是低效和富有挑戰(zhàn)性的，特別是因?yàn)槟繕?biāo)細(xì)胞與不需要的顆粒物常常具有相同的大小。目標(biāo)細(xì)胞同

5、時(shí)也非常脆弱和易受剪切力損傷，進(jìn)一步增加了細(xì)胞分離的復(fù)雜性。主要舉措應(yīng)對(duì)這些挑戰(zhàn)需要整個(gè)細(xì)胞制造產(chǎn)業(yè)藝術(shù)性的協(xié)同合作來開發(fā)、優(yōu)化和應(yīng)用先進(jìn)的細(xì)胞處理科技和技術(shù)。為了實(shí)現(xiàn)大規(guī)模，低成本，可持續(xù)生產(chǎn)高質(zhì)量細(xì)胞，細(xì)胞制造產(chǎn)業(yè)必須在以下關(guān)鍵細(xì)胞加工方面加強(qiáng)合作，包括：篩選方法；培養(yǎng)基優(yōu)化；細(xì)胞擴(kuò)增設(shè)備；細(xì)胞擴(kuò)增、修飾和分化方法；和分離技術(shù)。這些建議的具體措施在圖中有詳細(xì)描述，分為近期（20-2018中期（20-202和長(zhǎng)期（2022-2025）三個(gè)時(shí)間段。art.篩選方法基于細(xì)胞的治療，設(shè)備或診斷技術(shù)的治療效果決定于針對(duì)特定醫(yī)療產(chǎn)品所需特定屬性細(xì)胞的選擇性。目前選擇合適活細(xì)胞的處理方式是通過大量的反復(fù)

6、試驗(yàn)；因此需要開發(fā)和優(yōu)化高度自動(dòng)化畫的細(xì)胞篩選方法。先進(jìn)的供體和細(xì)胞篩選方法，包括使用更復(fù)雜的監(jiān)測(cè)和成像技術(shù)，將會(huì)使得細(xì)胞制造商更快速，高效，準(zhǔn)確地評(píng)估和選擇細(xì)胞。art.2培養(yǎng)基優(yōu)化為了制造批次規(guī)模達(dá)到億萬級(jí)的細(xì)胞，細(xì)胞制造產(chǎn)業(yè)必須發(fā)展非線性培養(yǎng)方式，如，懸浮培養(yǎng)方式來優(yōu)化細(xì)胞生產(chǎn)能力。不需要基底，微載體或飼養(yǎng)細(xì)胞的培養(yǎng)系統(tǒng)可以更好地利用空間，減少勞動(dòng)力需求，并根除目前細(xì)胞制造的一個(gè)主要成本驅(qū)動(dòng)因素。細(xì)胞制造產(chǎn)業(yè)有機(jī)會(huì)開發(fā)更多化學(xué)成分確定的培養(yǎng)基替代產(chǎn)品用來降低動(dòng)物來源污染的風(fēng)險(xiǎn)，并克服臨床級(jí)血清來源的限制性。先進(jìn)的細(xì)胞培養(yǎng)方式，便宜且可靠地培養(yǎng)基，以及更精確的細(xì)胞傳代方法可以降低細(xì)胞處理成

7、本并同時(shí)最大化細(xì)胞產(chǎn)量和最優(yōu)化細(xì)胞質(zhì)量。art.3細(xì)胞擴(kuò)增設(shè)備為了支持制造各類新細(xì)胞產(chǎn)品的技術(shù)，下一代細(xì)胞擴(kuò)增設(shè)備必須能夠適應(yīng)不同批次規(guī)模和并行處理不同類型細(xì)胞的能力。具有并行處理能力的（seedtrains）馴化種子和生物反應(yīng)器可以提高生產(chǎn)通量并處理時(shí)間，同時(shí)分?jǐn)偝杀局炼鄠€(gè)批次。先進(jìn)的細(xì)胞擴(kuò)增技術(shù)必須滿足高度自動(dòng)化，在封閉系統(tǒng)中進(jìn)行，并可被監(jiān)測(cè)和維持在確定理化水平上的要求。從而生產(chǎn)出具有批次間可比較性的細(xì)胞產(chǎn)品。封閉系統(tǒng)，并行處理能力和增強(qiáng)的自動(dòng)化水平是減少錯(cuò)誤和人與細(xì)胞產(chǎn)品相互作用導(dǎo)致污染的關(guān)鍵。art.4ffl胞擴(kuò)增、修飾和分化方法為了更精確和有效的區(qū)分特定細(xì)胞產(chǎn)品所需的具有目標(biāo)特性的細(xì)

8、胞，細(xì)胞制造產(chǎn)業(yè)必須增強(qiáng)其對(duì)細(xì)胞生物學(xué)的理解。更深入的了解為什么細(xì)胞會(huì)如止一包括，他們?nèi)绾雾憫?yīng)培養(yǎng)基和環(huán)境條件，他們加倍的速度，以及培養(yǎng)他們的最佳時(shí)I這些將增強(qiáng)對(duì)細(xì)胞屬性的控制。工業(yè)上還可以通過加速細(xì)胞分化的生物學(xué)速度來減少處理時(shí)間并增加生產(chǎn)能力（例如，運(yùn)用復(fù)雜的細(xì)胞培養(yǎng)和表觀遺傳學(xué)技術(shù)）或者減少細(xì)胞加倍時(shí)間。提高擴(kuò)增和分化的速度對(duì)于多能干細(xì)胞是特別重要的，應(yīng)為其通常需要幾個(gè)月時(shí)間來完成增殖和分化，這嚴(yán)重放慢了細(xì)胞產(chǎn)品進(jìn)入市場(chǎng)的時(shí)間和數(shù)量。art.分離技術(shù)將活細(xì)胞從培養(yǎng)基，無活性產(chǎn)物，為分化的細(xì)胞和其他顆粒物中分離，對(duì)細(xì)胞產(chǎn)品的純度，質(zhì)量和安全性至關(guān)重要。開發(fā)先進(jìn)的細(xì)胞分離技術(shù)，特別是下一代多

9、參數(shù)密閉系統(tǒng)，是減少細(xì)胞接種和細(xì)胞擴(kuò)增所需勞動(dòng)力的需求（例如，從微載體和基底上分離）。更高效的分離技術(shù)還可以減少細(xì)胞處理過程所需的培養(yǎng)基和空間需求。圖4,細(xì)胞處理主要建議（1）圖4,細(xì)胞處理主要建議（2）口圖4,細(xì)胞處理主要建議（3）圖4,細(xì)胞處理主要建議（4）口英文原文Cellprocessingthegrowthofcellsinanartificialenvironmentoutsideofthehumanbodyisadefiningpartofthecellmanufacturingprocess.Eachparameterofcellculturing,includingtheve

10、ssel,media,nutrients,andphysicochemicalenvironment,influencesthepropertiesthatcellsrequiretobeefectivetherapeuticallyThedevelopmentofnewcell-basedmedicalproductswilldemandcellprocessingtechnologiesandtechniquescapableofmanufacturingcellsingreaterquantitiesatgreaterspeeds.However,thematerials,space,l

11、abor,andtimerequirementsofcurrentmethodswillpreventexistingprocessesfrommeetingthisgrowingdemand.Toenablelarge-scalemanufacturingofhigh-qualitycellsinthenext10years,thecellmanufacturingcommunitymustworktogethertodevelop,optimize,andimplementmorecost-efectiveandeficientcellprocessingtechnologiesandte

12、chniques.Approachesthatcanalsoincreasetheconsistencyandreliabilityofcellprocessingcouldenhancethequalityandeficacyofcell-basedproducts.CurrentChallengesTorealizelarge-scale,cost-efective,reproduciblemanufacturingofhigh-qualitycells,thecellmanufacturingcommunitymustworktoovercomethecellprocessingchal

13、lengesthatfollow.art.1inearnatureofexistingcultureplatformsMostcurrentcultureplatforms(e.g.,planarculturesystemsorfeedercells)arelinearandrequireadditionalsurfaceareatomanufacturecellsingreaterquantities.Theresultingspaceconstraints,coupledwiththesubstantialculturemediaandlaborrequirementsofcurrents

14、ystems,renderexistingculturingapproacheseconomicallyimpracticalforcommercial-scalemanufacturing.Additionally,itisdificulttocontrolnutrientgradientsacrosstheselinearsystemstoachieveconsistentcellcharacteristicsandqualityart.Increasedpotentialforcontaminationinopen-culturesettingsInconventionalresearc

15、hsettings,thecurrentprocessofdividingcellculturestoenablefurtherproliferationiscarriedoutinopensystemswherevesselsareexposedtothesurroundingenvironment.Everyopeningofaculturevesselposesthepotentialforcontaminationfrommolds,yeasts,viruses,mycoplasma,andothercelllines.Thecaretakentomaintainsterileenvi

16、ronmentstoreducethisriskistime-andlabor-intensive,necessitatingtheneedforinnovativeclosed-systemtechnologiesthataresealedfromtheexternalenvironment.art.DifcultyunderstandingcellcomplexityThecellmanufacturingcommunitylackssuficientunderstandingofthebiologyandemergentpropertiesofhumancells,particularl

17、ystemcellsandimmunecells.Understandingofhumancellsisfurthercomplicatedbythefactthatcellsproducedthroughcellmanufacturingdiferfromthoseinthebody.Becauseoftheinherentcomplexityofcells,itisdificulttodefinetheneededpropertiesorthemodeofactionforcellstobeusefultherapeuticallyorthecultureenvironmentsanddi

18、ferentiationprocessesnecessarytomanufacturecellswiththeseproperties.art.Highcost,limitedsupply,andinconsistencyofrawmaterialsCulturemediaincludinggrowthfactors,nutrients,andreagentsareofenthemostexpensivepartofthecellmanufacturingprocess.Thehighcost,aswellasthelimitedsupplyandshelflifeofsomerawmater

19、ials,willpreventexistingcultureplatformsfrommeetingincreasedcellproductiondemand.Anadditionalcomplicationisthatsomemediacomponents,particularlyanimalserums,havebatch-to-batchvariability,whichcanreducecellpropertyconsistencyandevencontaminatecellproducts.art.InefciencyofcellseparationmethodsInmanycur

20、rentcellprocessingsystems,cellsmustberemovedfromthesurfaceofculturingvesselsandseparatedintonewvesselswithfreshgrowthmediumforfurtherpropagation.Whenpreparingcellsfordistribution,cellmustalsobeseparatedfromculturemedia,undiferentiatedcells,andotherparticulatestoachievethecellpurityneededfortheendpro

21、duct.Currentseparationprocessesareineficientandchallenging,particularlybecausedesiredcellsareofenthesamesizeasunwantedparticles.Desiredcellsarealsofragileandsubjecttoshear,furtheraddingtothecomplexityofcellseparation.KeyInitiativesAddressingthesechallengeswillrequirecoordinatedefortsacrossthecellman

22、ufacturingcommunitytodevelop,optimize,andimplementadvancedcellprocessingtechnologiesandtechniques.Torealizethepotentialoflarge-scale,cost-efective,reproduciblemanufacturingofhigh-qualitycells,thecellmanufacturingcommunitymustcollaborateonthefollowingkeycellprocessinginitiatives:ScreeningandSelection

23、Methods;CultureMediaAdvances;CellExpansionEquipment;CellExpansion,Modification,andDiferentiationMethods;andSeparationTechniques.ActivitiesforeachoftheseinitiativesareprovidedinFigure4,dividedintonear-term(2016-2018),mid-term(2019-2021),andlong-term(2022-2025)timeframes.art.1ScreeningandSelectionMeth

24、odsThetherapeuticefectivenessofcell-basedtreatments,devices,ordiagnostictechnologiesdemandstheselectionofcellswiththedesiredpropertiesforthatspecificmedicalproduct.Currentprocessesforselectingviable,suitablecellscaninvolveextensivetrialanderror,necessitatingthedevelopmentandoptimizationofhighlyautom

25、atedcellscreeningandselectionmethods.Advancedapproachestodonorandcellscreening,includingtheuseofmoresophisticatedassaysandimagingtechnologies,couldenablecellmanufacturerstomorequickly,eficiently,andaccuratelyassessandselectcells.art.2CultureMediaAdvancesTomanufacturelotsizeswithtrillionsofcells,thec

26、ellmanufacturingcommunitymustadvancenon-linearcultureformats,suchassuspensioncultures,thatoptimizecellularproductivity.Withouttheneedforsubstrates,microcarriers,orfeedercells,suchculturesystemscouldbetterutilizespace,reducelaborrequirements,andeliminatewhatiscurrentlyoneoftheprimarycostdriversofcell

27、manufacturing.Thecellmanufacturingcommunityhastheopportunitytodevelopmorechemicallydefinedmediaalternativestoreducetheriskofcontaminationfromanimalmaterialsandovercomethelimitedavailabilityofclinical-gradesera.Advancedcellculturingformats,lower-costandmorereliablemedia,andmoreprecisecellpassagingcou

28、ldreducecellprocessingcostswhilealsomaximizingcellyieldandquality.art.CellExpansionEquipmentTosupportthemanufacturingofavarietyofnewcell-basedtechnologies,next-generationcellexpansionequipmentmustbeabletoaccommodatevaryinglotsizesandparallelprocessingofdiferentcelltypes.Seedtrainsandbioreactorswithp

29、arallelprocessingcapabilitiescouldincreasemanufacturingthroughputandshortenprocessingtimewhiledistributingcostovermultiplebatches.Advancedcellexpansionmustalsobehighlyautomatedandconductedinclosedsystemsthatcanbemonitoredandmaintainedatdefinedphysiochemicallevels,resultingincultureswithcomparablecha

30、racteristicsfrombatchtobatch.Closed-system,parallelprocessingwithincreasedautomationisalsocriticaltominimizeerrorandcontaminationfromhumaninteractionwithcellproducts.art.4CellExpansion,Modifcation,andDiferentiationMethodsTomoreaccuratelyandeficientlydiferentiatecellswiththedesiredcharacteristicsfora

31、cell-basedmedicalproduct,thecellmanufacturingcommunitymustincreaseitsunderstandingofcellbiology.Increasedunderstandingofwhycellsdowhattheydoincludinghowtheyrespondtomediaandenvironmentalconditions,thespeedatwhichtheydouble,andtheoptimumtimetoculturethemwillenableincreasedcontrolofcellproperties.Theindustrycouldalsoreduceprocessingtimes