Læknablaðið - 15.05.1998, Page 9

LÆKNABLAÐIÐ 1998; 84
373
Stofnfrumugræðlingar
blóðmyndandi vefs
Kristbjörn Orri Guömundsson’1, Leifur Þorsteinsson11, Ásgeir Haraldsson2’, Sveinn Guðmundsson’’
Guðmundsson KO, Þorsteinsson L, Haraldsson Á,
Guðmundsson S
Hematopoietic stem cell grafts
Læknablaðið 1998; 84: 373-9
Introduction: The number of stem cell transplanta-
tions have greatly increased in the treatment of pri-
mary bone marrow disorders and as a rescue therapy
following high-dose chemoradiotherapy for various
malignancies. Hematopoietic stem cells can be
mobilized from bone marrow by hematopoietic
growth factors enabling their sampling from perip-
heral blood. Peripheral blood stem cell transplanta-
tion is gradually replacing autologous bone marrow
transplantation and is increasingly used in allogeneic
settings. Transplantation using umbilical cord blood
derived stem cells are also well described.
Objective: To standardize the in vitro methods nec-
essary for the evaluation of stem cell grafts using
umbilical cord blood mononuclear cells.
Material and methods: Mononuclear cells (MNC)
were isolated from 57 umbilical cord blood samples.
The proportion of hematopoietic stem cells (CD34+)
was determined by flow cytometry. The number of
clonogenic cells (BFU-E, CFU-GM) was determined
by culturing MNC in methylcellulose and agar. The
number of clonogenic cells was compared before and
after freezing in liquid nitrogen.
Results: The mean volume of collected cord blood
was 43.8 ml and the number of MNC’s was
102.7XI011 cells of which 0.93% were CD34+. The
number of CFU-GM was 238/105 MNC and BFU-E
506/105 MNC. After freezing and thawing the MNC,
the viability was 94.9% and the number of clonogen-
Frá "Blóðbankanum,z|Barnaspítala Hringsins Landspltal-
anum. Fyrirspurnir, bréfaskipti: Kristbjörn Orri Guðmunds-
son, Blóðbankanum, 101 Reykjavík. Sími: 560 2020; bréf-
sími: 562 3051; netfang: kristbj@rsp.is
Lykilorð: stofnfruma, CD34, któnógenísk ræktun.
ic cells was slightly decreased when compared to
pre-freezing values, the difference being not statist-
ically significant. The purity of CD34+ cells after
selection with magnetic beads was over 95%.
Conclusion: In vitro methods, neccessary for evalu-
ation of hematopoietic stem cell grafts, have been
standardized using umbilical cord blood derived
stem cells.
Keywords: stem cell, CD34, clonogenic culture.
Ágrip
Inngangur: Mögulegt er að auka fjölda
blóðmyndandi stofnfrumna í blóðrás og ein-
angra þær þaðan. Notkun þeirra í ígræðslum
hefur aukist mikið undanfarin ár sem þáttur í
meðferð illkynja blóðsjúkdóma og fastra æxla.
Igræðslur stofnfrumna úr blóðrás hafa að mestu
leyti komið í stað samgena (autologous) bein-
mergsígræðslna en eru einnig að ryðja sér til
rúms í stað ósamgena (allogeneic) beinmergs-
ígræðslna. Blóðmyndandi stofnfrumur er hægt
að einangra úr naflastrengsblóði og nota til
ígræðslu.
Markmið: Að staðla in vitro aðferðir sem
eru nauðsynlegar við ígræðslur blóðmyndandi
vefs og nota til þess einkjarna hvítfrumur úr
naflastrengsblóði.
Efniviður og aðferðir: Einkjarna hvítfrum-
ur (mononuclear cells, MNC) voru einangraðar
úr blóði 57 naflastrengja. Hlutfall CD34+
frumna var mælt með frumuflæðisjárgreiningu.
Mat á fjölda kólóníumyndandi frumna (CFU-
GM, BFU-E) var ákvarðað með ræktun ein-
kjarna hvítfrumna í metýlsellulósa- og agaræti.
Kólóníuvöxtur var borinn saman fyrir og eftir
frystingu í fljótandi köfnunarefni.
Niðurstöður: Meðalrúmmál naflastrengs-
blóðs sem safnaðist var 43,8 ml. Fjöldi ein-