Gupta Namita, Chelluri Lakshmi Kiran, Ratnakar Kamaraju Suguna, Ravindhranath K, Vasantha A
Transplant Immunology and Stem cell Laboratory, Global Hospitals, Hyderabad, India.
Asian J Transfus Sci. 2008 Jul;2(2):69-80. doi: 10.4103/0973-6247.42694.
Concentrations of O(2) and CO(2) in the fetal circulation differ to that in maternal blood. Previous studies done in algae demonstrate the functional role of Rh antigen as CO(2) transporter. As a preliminary study, it was the aim of this project to compare the expression of Rh polypeptides on cord and adult red blood cell progenitors during ex vivo proliferation and differentiation of CD34(+) cells during erythropoeisis.
CD34 positive hematopoeitic progenitor cells were isolated from umbilical cord blood and adult peripheral blood using an immunomagnetic system and cultured in serum free medium containing erythropoietin in order to compel them along the erythroid lineage. Cultured cells were analyzed for cell surface marker expression by flow cytometry, using monoclonal antibodies to RhAG, Glycophorin A, Rh polypeptides, CD47 and Band 3. Cytospin analysis was also done to study the morphology of cultured cells.
The appearance of cell surface markers analyzed on different days of culture varied slightly between samples. There was no evidence to suggest that RhAG, GPA, CD47 and Band 3 expression was any different between adult and cord derived cells. Nevertheless, the results of Rh antigenic expression suggest a reasonable difference between the two groups with adult sample derived cells showing higher and earlier expression than cord blood derived cells. These preliminary findings require further investigation.
Comparing the expression of cell surface markers especially Rh polypeptides between adult and cord blood derived erythroid progenitors might assist in discerning their functions and could be valuable in the study of erythropoeisis.
胎儿循环中氧气(O₂)和二氧化碳(CO₂)的浓度与母体血液中的不同。先前在藻类中进行的研究证明了Rh抗原作为CO₂转运体的功能作用。作为一项初步研究,本项目的目的是比较在红细胞生成过程中CD34⁺细胞体外增殖和分化期间脐带血和成人红细胞祖细胞上Rh多肽的表达。
使用免疫磁珠系统从脐带血和成人外周血中分离出CD34阳性造血祖细胞,并在含有促红细胞生成素的无血清培养基中培养,以使它们沿着红系谱系分化。使用针对RhAG、血型糖蛋白A、Rh多肽、CD47和带3的单克隆抗体,通过流式细胞术分析培养细胞的细胞表面标志物表达。还进行了细胞涂片分析以研究培养细胞的形态。
在不同培养天数分析的细胞表面标志物的出现情况在样本之间略有不同。没有证据表明成人来源细胞和脐带血来源细胞之间的RhAG、GPA、CD47和带3表达有任何差异。然而,Rh抗原表达的结果表明两组之间存在合理差异,成人来源细胞的表达高于且早于脐带血来源细胞。这些初步发现需要进一步研究。
比较成人和脐带血来源的红系祖细胞之间细胞表面标志物尤其是Rh多肽的表达,可能有助于识别它们的功能,并且在红细胞生成的研究中可能具有重要价值。