Differential effects of p40<i><sup>ABL/BCR</sup></i>, p96<i><sup>ABL/BCR</sup></i> and p185<i><sup>BCR/ABL</sup></i> on the B cell commitment of murine HSCs.

Abstract

<p>(A) Experimental strategy for studying the influence of the reciprocal t(9;22) fusion proteins on the B cell commitment of murine HSCs. Sca1<sup>+</sup>/lin<sup>−</sup> BM cells were infected with the indicated retroviruses and plated in semi-solid medium supplemented with the indicated growth factors for determination of the serial replating potential. Cells from the first plating (I) round and cells plated in liquid culture supplemented with the indicated growth factors were examined for the expression of differentiation-specific surface markers. (B) Long-term serial replating. Sca1<sup>+</sup>/lin<sup>−</sup> cells were infected with the indicated retroviruses and plated into methyl-cellulose medium supplemented with the indicated growth factors to assess primary colony formation. Colony numbers were counted on days 8–10. Cells were then harvested and serially replated. Colonies were counted on days 8–10 for each replating. I-IV - number of the plating round. (C) Expression of differentiation-specific surface markers. (D) Colony morphology of the second plating. Type A (compact colonies), type B (a dense center surrounded by a halo of migrating cells) and type C (diffuse colonies with mobile differentiating cells) colonies were distinguished. p185<i><sup>BCR/ABL</sup></i> exhibited a high number of viable cells that were not organized into colonies. (E) Determination of the maturation stage of B220<sup>+</sup> lymphocytes by CD43 expression analysis. CD43 positivity is characteristic of immature pro-B cells.</p

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