Plots are fractionated by each ancestor type (SLAM-HSCs, ST-HSCs, and multi-potent progenitors [MPPs]) and cocktail (with and without IL-3 + IL-6, indicated by + and -, respectively). (A) Simultaneous visualization of family membership, generation number, and cell type of offspring from initially seeded ancestors harvested at two time points (24 and 48 hr). Each row presents the offspring from a single ancestor. Columns identify the generation number of each recovered cell, with their phenotypic cell type indicated by color coding. Rows are sorted in increasing order of the difference between maximum and minimum generations in each ancestor’s family (generational range). (B) To test the differentiation pattern of our cell phenotypes after culture, sorted cell types as shown in Figure 1 were cultured in methylcellulose cultures and evaluated for colony forming units (CFUs) formation for megakaryocytic (MK), granulocytic/monocytic (GM), erythroid burst-forming (BFU-E), or mixed colony formation of all types (CFU-MK, CFU-GM, and BFU-E) with or without MK. Late MP did not produce any colonies in methylcellulose culture (C) The percentages of MK, CFU-MK, and mixed colony formation are indicated from the culture in serum-free fibrin clot. GMP did not produce any colonies as expected. (D) Percentage of each recovered cell type. (E) Distribution of the maximum generation per family, as indicated by color coding. (F) Proportions of recovered cell types for ancestors that have not yet divided. (G) For ancestors that have divided only once and for whom two offspring are recovered, percentage having symmetric and asymmetric fates with and without differentiation. (H) Percentage of families with each generational range. The 48 hr data (red bars) is compared to a mathematical model parameterized by a single coefficient, ρ, which encodes the correlation in whether cells in the same generation within a family divide or cease to divide (Materials and methods). Shown is prediction for both the maximum likelihood best-fit value of ρ (value and blue/white bars) and, as a reference, a range of other values of ρ (solid blue bars). For (D–H), error bars indicate 95% confidence intervals calculated via basic bootstrap (Materials and methods). Sample sizes and p-values (from permutation tests, see Materials and methods) from the panels can be found in Figure 2—source data 1 and 2.