来源于骨髓、外周血与脐带血的间充质干细胞生物学特性比较
{;:死震基础医学
-¨目纰 织 jm !研 究 ’j怖』术康 复 彩 73 誉 彩 4 5 助‘ 2009 — 1 1 — 05 出版
J ournalofC linicalR ehabilitative T issue E ngm eefingR esearch N ovem ber5 , 2009 V 0 1. 13 , N o . 4 5
来源于骨髓、 外周血与脐带血的间充质干细胞生物学特性比较术
黄友章’, 沈建 良’, 寓立 众’, 尹 文杰 ’, 刘 毅 ’, 成 海。, 郑培浩’, 岑 坚 ’
C o...
{;:死震基础医学
-¨目纰 织 jm !研 究 ’j怖』术康 复 彩 73 誉 彩 4 5 助‘ 2009 — 1 1 — 05 出版
J ournalofC linicalR ehabilitative T issue E ngm eefingR esearch N ovem ber5 , 2009 V 0 1. 13 , N o . 4 5
来源于骨髓、 外周血与脐带血的间充质干细胞生物学特性比较术
黄友章’, 沈建 良’, 寓立 众’, 尹 文杰 ’, 刘 毅 ’, 成 海。, 郑培浩’, 岑 坚 ’
C om parison ofbio logicalcharacteristics ofm esenchym alstem cells derived fro m bone m arrow ,
peripheralblood and cord blood
H uangY ou- zhang
’
, S hen J ian— liang
’
, G ong L i? zhong
’
, Y in W en- jie’, L iu Y i’, C heng H al。. Z heng P ei. hao’. C en J ian’
’D epartm entof
H em atology,
‘D epartm entofB Iood
T ransfusion . Navy
G eneralH ospitalof
C hinese P L A . B eijing
1 0004 8 C hina
H uangY ou— zhang,
A ssociate chief
technician。
D epartm entof
H em atology, Navy
G eneraI H ospitalof
C hinese P L A , B eijing
10004 8 C hina
huangyouzhang08@
yahoo corn cn
S upported by:the
S cient砺c R esearch
P lan ofM inistryof
H ealthandNavy
L ogistics D epartm ent.
No 200828203 ‘
R eceived:2009 — 06- 09
A ccepted:2009 — 09- 01
解 放 军 海 军 总 医
院 , ’ 血 液 科 ,
。 输 血 科 , 北 京市
1 0004 8
黄友 章 , 男 , 195 6
年 生 , 湖 南省长 沙
市 人 , 汉 族 , 1992
年 中 国 科 技 大 学
毕 业 , 副 主 任 技
师 , 主要 从 事血 液
学 方 面 的 研 究 。
huangyO uznang
08@ yahoo.
C O m C n
海 军 后 勤 部 卫 生
部 科 研 计 划 课 题
(200828203)’
?}‘罔 分娄 呼:R 394 2
文献 标识 码:B
文 章编 号:1673 - 8225
(2009)4 5-08966- 05
收 稿 日期 :2009 . 06- 09
修 回 日期 :2009 — 09 - 01
f200906090'4 ,
Z S ‘ Q )
8966
A bstract
B A C K G R O U N D :M esenchym alstem cells (M S C s)existin hum an tissues P resently, cellsource is single;culture m ethodhas
greatdiffe rences;obtained results are notconsistent T hus, itcannotverfythatisolated andcultured cells are jdenticaJ cells. w hich
is difficultco com pare .
O B J E C T IV E :T o com pare the biologicalfeatures ofM S C s derived form bone m arrow (B M ), perpheralblood (P B )andcord blood
(C B )underin vitro culture conditions.
D E S IG N , T IM E A N D S E T T IN G :T he cytologicalin vitro controlledstudyw as perfo rm ed atthe D epartm entofH em atology。 N avy
G eneralH ospitalofC hinese P L A from J une 2007 to D ecem be r2008.
M A T E R IA L S :A totalof10 donors ofhem opoietic stem celltransplantatio n atthe D epartm entofH em atology N avyG eneral
H ospitalofC hinese P L A w ere selected. M B and P B cells w ere obtainedfrom the sam e donor, and cellvolum es w ere respectively
20 m L and 2 m L _C B cells (30 m L )w ere obtainedfrom healthyprim ipara atthe D epartm entofO bstetrics, N avy G enerafH ospitalof
C hinese P L A .
M E T H O D S : M S C s w ere obtainedfrO m B M , P B and C B by P ercolIdensitygradient+ adherence m ethod. andthen incubated in
D M E M /F 12 m edium containing 10% fetaIbovine serum W hen 80% 90% confluency cells w ere digested in trypsin. E D T A and
m ade into 5 × 10。/L celIsuspension as P 0 A bove— describedoperation w as perform ed as P 1 andthe restm ay be deduced by
analogyas P 2
一 P s.
M A lN 0 U T C O M E M E A S U R E S :T he follow ing param eters w ere m easured:cellgrow thm orphology;results ofW right. G iem sa
staining;results ofcytochem istry:celI proliferation am ount:cellsurface m arkers usingflow cytom etry .
R E S U L T S :T im e ofadherence. tim e to 50% confluencyandtim e to 80% confluencyofB M S C s w ere earliercom arpedw iththe
P B M S C s and U C M S C s. A dherentcells from B M grew in w hirpool— like type . w hile C B and P B did nolat5 7 days M ajorityof
aderentcells from B M w ere fibroblast— like cells. and sm alIparts w ere endothelioidcells. A derenlcells frO m P B and C B atthe fifth
generation contained m ore endothelioidcells and m ononuclear and m acrophage— like cells besides fibroblast— like cells. P A S stain,
S udan blackB stain, alkaline phosphatase (A K P )stainingofadherentcells from B M , P B andC B w ere negative from P 1 Io P 5
C om paredw ith P 0 cells, num berofB M M S C s lillP 5 w as significantlym ore in P B M S C s and U C M S C s (P ( 0. 05 ) P ositive rates of
C D 29 , C D 4 4 . C D 90, C D 7 1 , C D l05 , C D l66 and H L A — A B C w ere 55 9% 一 92 . 8% atP 0 to P 5 , bul ≤ 6% foIlow ing B M M S C s w ere
incubated:19 17% 33 .4 % alP 0 to P 5 , but ≤ 1 0% follow ingP B M S C s w ere incubated;35 .4 % 93 _2% alP 0 to P 5 , bul ≤ 20%
follow ingC B M S C s w ere incubated. P ositive rates ofC D 34 . C D 4 5 and H L A — D R w ere low in B M - . P B — andC B — M S C s. P ositive rates
O fC D l4 andC D 3 1 w ere low in B M M S C s;12 1 % 283% in P B M S C s, and8. 1 % 21 3% in C B M S C s
C O N C L U S IO N :M S C s can be attainedfrO m B M , P B andC B Q uantities ofM S C s form B M are the highest. w ithsingle com ponent.
foIlow ed by C B M S C s and P B M S C s. w ith m ultiple com ponents
H uangY Z , S hen J L 。 G ongL Z , Y in W J , L iu Y , C hengH , Z hengP H , C en J . C om parison ofbiologicalcharacteristics ofm esenchym a
stem cells derivedfrom bone m arrow . peripheralblood andcord blood. Z hongguo Z . uzhiG ongchengY anjiu yu L inchuang K angfu
2009 ;1 3(4 5 ):8966- 8970 . [http://w w w . crterca http://en zglckf. com 】
摘 要
背景 : 问充质 干 细胞存在 于人体多种组 织 , 目前研 究报道 所用细胞来源 单 一 , 培养