A preliminary study on the culture of single hematopoietic stem cell / 中国医学科学院学报
Acta Academiae Medicinae Sinicae
; (6): 19-24, 2012.
Article
en Zh
| WPRIM
| ID: wpr-352956
Biblioteca responsable:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the biological behavior including survival and proliferation of CD34 + CD38--Lin--cells when they are cultured at single cell level.</p><p><b>METHODS</b>Purified umbilical cord blood CD34 + CD38--Lin--cells were separated at single cell level in 96-well plates using flow cytometry for four groups: control group (CD34 + CD38--Lin--cell plus stem cell medium) , Shh group (CD34 + CD38--Lin--cell plus stem cell medium and Shh), BMP-4 group (CD34 + CD38--Lin--cell plus stem cell medium and BMP-4), Jagged-1 group (CD34 + CD38--Lin--cell plus stem cell medium and Jagged-1). Methylcellulose medium was used in the colony-forming experiment which was also in four groups as previously. The number of cells and colony-forming units in each well for the four groups was evaluated at different time points (day 1, 3, 7) with fluorescence microscopy counting method.</p><p><b>RESULTS</b>Division of single cell was observed to be amplified in all of these groups from day 3. And meanwhile, after 1-week culture, the survival rates for the treated groups were all higher than the control group (Jagged-1 group > BMP-4 group > Shh group > control), while the cell number in each well was also highest in the Jagged-1 group (Jagged-1 group > BMP-4 group > control). The number of wells with a cell number of zero was significantly fewer in all treated groups (especially the Jagged-1 group) than in the control group; meanwhile, the number of wells with a cell number higher than 17 was evidently higher in all the treated groups (especially the BMP-4 group) more than controls. Colony-forming units for erythroid (BFU-E), granulocyte (CFU-G), macrophage (CFU-M), and granulocyte macrophage (CFU-GM) were observed for all of these experimental groups, and there was no significant difference between the four experimental groups.</p><p><b>CONCLUSIONS</b>CD34 + CD38 - Lin - cell can achieve the survival, self-renewal and proliferation when cultured at single cell level, and the adding of Shh, BMP-4, and Jagged-1 can enhance such capabilities. However, CD34 + CD38 - Lin - cell can only maintain cell totipotency in its niche.</p>
Texto completo:
1
Base de datos:
WPRIM
Asunto principal:
Proteínas de Unión al Calcio
/
Células Madre Hematopoyéticas
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Supervivencia Celular
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Células Cultivadas
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Química
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Ensayo de Unidades Formadoras de Colonias
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Técnicas de Cultivo de Célula
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Antígenos CD34
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Medios de Cultivo
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Biología Celular
Límite:
Humans
Idioma:
Zh
Revista:
Acta Academiae Medicinae Sinicae
Año:
2012
Tipo del documento:
Article