Sun Ai-hong, Feng Si-zhou, Liu Bin, Tang Feng-wu, Feng Yi, Lu Shi-hong, Han Zhong-chao
State Key Laboratory of Experimental Hematology and Center of Hematopoietical Stem Transplantation, Institute of Hematology & Blood Diseases Hospital, CAMS & PUMC, Tianjin 300020, China.
Zhonghua Xue Ye Xue Za Zhi. 2005 May;26(5):261-4.
To evaluate the hematopoietic reconstitution of implanted NOD/SCID mice, after intra-bone marrow cavity injection (iBM) of human umbilical cord blood (CB) mononuclear cells (MNCs).
24 female NOD/SCID mice were divided into different MNCs dosage iBM groups (3 x 10(6), 1 x 10(7), 3 x 10(7) cells), tail vein intravenous injection (iTV) group (3 x 10(7) cells) and control group (iBM of medium only). CB MNCs sorted by Ficoll-Hypaque were transplanted into left tibia bone marrow cavity of 6- to 8-week-old NOD/SCID mice, which were anesthetized and sublethally irradiated (270 cGy (137)Cs-gamma irradiation). The distribution of injected CB MNCs in noninjected right tibia of the same implanted mice was observed 24 hours after iBM. The establishment of hematopoiesis and the survival of mice were observed. BM cell surface CD marker expressions, dye Dil-CM tracing and human beta-actin from implanted mice were assessed 8 weeks after iBM or iTV.
Dil-CM marker could be detected on BM cells from noninjected right tibia 24 hours after iBM. Fourteen engrafted mice survived at the end of our study. Among them two, four and five were of iBM-1, iBM-2 and iBM-3 groups respectively, and one of control group and two of iTV group. White blood cell reconstitution was better in iBM mice than in iTV and control mice. There were human markers including CD45, Dil-CM and beta-actin DNA in the marrow cells from the human CB MNC engrafted mice.
The preliminary results showed that hematopoiesis reconstitution by iBM was significantly better than iTV.
评估经骨髓腔内注射（iBM）人脐带血（CB）单个核细胞（MNCs）后，植入的NOD/SCID小鼠的造血重建情况。
将24只雌性NOD/SCID小鼠分为不同MNCs剂量的iBM组（3×10⁶、1×10⁷、3×10⁷个细胞）、尾静脉注射（iTV）组（3×10⁷个细胞）和对照组（仅注射培养基的iBM组）。通过Ficoll-Hypaque分离的CB MNCs被移植到6至8周龄经麻醉并接受亚致死剂量照射（270 cGy ¹³⁷Cs-γ射线照射）的NOD/SCID小鼠的左胫骨骨髓腔内。iBM后24小时观察注射的CB MNCs在同一植入小鼠未注射右胫骨中的分布情况。观察造血的建立和小鼠的存活情况。在iBM或iTV后8周评估植入小鼠的骨髓细胞表面CD标志物表达、Dil-CM染料示踪和人β-肌动蛋白。
iBM后24小时，在未注射右胫骨的骨髓细胞上可检测到Dil-CM标志物。在我们研究结束时，有14只移植成功的小鼠存活。其中，iBM-1组有2只，iBM-2组有4只，iBM-3组有5只，对照组有1只，iTV组有2只。iBM小鼠的白细胞重建情况优于iTV和对照组小鼠。在人CB MNC植入小鼠的骨髓细胞中存在包括CD45、Dil-CM和β-肌动蛋白DNA在内的人标志物。
初步结果表明，iBM的造血重建明显优于iTV。
