【精品论文】体外扩增脐血间充质干细胞的生物学特性和诱导分化能力的研究

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1、论著体外扩增脐血间充质干细胞的生物学特性和诱导分化能力的研究鞠秀i'jij黄志伟时庆侯怀水段春红【摘要】目的从脐血中分离单个核细胞(MNCs),体外培养扩増间充质干细胞(MSCs)并研究其生物学特性和诱导分化潜能。方法取35份足月顺产新生儿脐血，密度梯度离心法分离出其中的MNCs,采用含胎1-.lfD.iW的DV1BV1培养基体外培养扩增MSCSo显微锐下观察MSCs的形态、细胞化学染色漩式细胞仪测定MSCs的细胞免疫表型，体外诱导分化实验检测MSCs分化成骨细胞、脂肪细胞和神经细胞的能力。结果从12份

2、脐血中可培养出MSCs,形态上与从其他來源的MSCs类似，可传至20代而无形态上的变化。细胞化学染色示碱性磷酸酶(ALP)阴性，非特异性酯酶一

3、丁酸荼酚酯酚(alpha-naphtholbutyricacidesterase,NBE)阳性。其表达和CDi®,待別是人类MSCs标记SH・2和SH・3阳性，而CD；、CDi4、a)i9、CD34和CD"阴性，说明它们并非来口造血细胞。这些MSCs在适当诱导分化剂的作用下，2周左右可以诱导分化形成骨细胞，20天左右可以诱导分化形成脂肪细胞。脐血MSCs预诱导1

4、2h后，胞体发生收缩，细胞边缘有细的突起。正式诱导5h后大多数细胞呈现典型的神经元样。结论胎儿脐血MNCs可分离培养出MSCSo这些MSCs具冇与其他來源MSCs类似的表型及分化潜能。【关键词】胎血；间质干细胞；细胞分化；细胞,培养的BiologicalcharacteristicsandhduceddfferentHtbnabilityofinvitroexpandedumbilicalcordbloodmesenchymalstancellsJUXhrliHUANGZhZeISHIQingfHOUH

5、uarshuiDUANChun-hongDepartnentofPediatries.QiluHospita/ofShandongUnivemity9Jinan250012,China【Abstract]ObjectiveMesenchymalstemcells(MSCs)fmmbonemamnvarecapableofdifferentiatingintocellsofdiffcrcnttissuelineagessuchasbone,cartilage,andadiposctissueandarct

6、hebestcandidatesfortissueengineeringItiswellacceptedthatimbilicalcordbkx)d(UCB)isasourceforhcmak)poicticstancellsHowever,conttovcrsyexistsasiowhcthcrUCBcontainsMSCsandcanserveasasourceofMSCsTlierefore,theainoftliisstudywasL)expbrethebiobgicalcharacterist

7、icsandinducingdifferentiationabilityofinvitn)expandedUCBMSCsMethodsUCBwascollectedonnoonaIfullteirndeliveryofinfontswithinfoimedconsent(n=35)obfciinedfiumthemothersMononuclearcells(MNCs)wereisolatedfianUCBbygravitycentrifugationandculturedwithDMEVIinclud

8、ing10%fetalbovineserumThemoiphobgywasobservedundermiciuscopeperdayCytochemicalstainingwascarriedoulandfbwcytometrywasusedIdexanincthesurfaceantigenphenotypeFifthpassageccllswcretransferredintoadifferentmediumandosteogenicdifferentiation,adipogenicdiffere

9、ntiation,andneurogenicdiffercntiationwereassessedResultsMSCscouldbeisolatedandcultuiedfictnMNCsofafewUCBjouncesThesecellsdisplayedfibroblast-likemoiphobgyTlieywithst>oclover20passageswithoutsignificantstmcturalchangesTlies