A new method of expanding cord blood cells

Details A new method of expanding cord blood cells A new method of expanding cord blood cells

C - Chemistry, Metallurgy

12

N

C12N 5/073 (2010.01) C12N 5/078 (2010.01) C12N 5/02 (2006.01)

Patent

CA 2562760

Based on previous evidence suggesting positive effects of fever on in vivo hematopoiesis, the effect of hyperthermia on the expansion and differentiation of megakaryocytes (MKs) in ex vivo cultures of CB CD34-enriched cells has now been tested. Cells were cultured at 37~C or 39~C for 14 days in cytokine conditions optimized for MK development, and analyzed periodically by microscopy, flow cytometry and colony assays. Compared to 37~C, cultures maintained at 39~C produced much more total cells (5X), MK progenitors (9X) and total MKs (7X), and showed accelerated (3-4 days) and enhanced MK maturation with increased yields of proplatelets and platelets (11.7X). The increased number of CD34+ cells and myeloid progenitors in the 39~C cultures also suggested a general stimulatory effect of hyperthermia on the expansion of more primitive stem/progenitor cells and of cells of other lineages.

En fonction de preuves précédentes suggérant des effets positifs de la fièvre sur l'hématopoïèse in vivo, l'effet de l'hyperthermie sur l'expansion et la différentiation de mégacaryocytes (MK) dans des cultures in vivo de cellules enrichies en CB CD34 a maintenant été testé. Des cellules ont été mises en culture à 37 ·C ou 39 ·C pendant 14 jours dans des conditions de cytokine optimisées pour le développement de MK, et analysées périodiquement par microscopie, cytométrie de flux et essais de colonie. Comparées à des cultures à 37 ·C, des cultures maintenues à 39 ·C ont produit significativement plus de cellules totales (5X), de progéniteur de MK (9X) et de MK au total (7X), et ont fait preuve d'une maturation de MK accélérée (3-4 jours) et renforcée avec des rendements accrus de plaquettes et proplaquettes (11,7X). L'augmentation de cellules CD34+ et de progéniteurs de myéloïde dans des cultures à 39 ·C suggère également un effet de stimulation général de l'hyperthermie sur l'expansion de cellules souches/progénitrices plus primitives et sur des cellules d'autres lignées.

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