American Society of Hematology

Figure 1.

$Optimizing flow cytometry–based measurements of drug sensitivity in MM patient samples ex vivo. (A) MM cell viability after 48 hours from 4 primary samples that were thawed and separated into fractions and cultured either without selection (MNCs) or with CD138 selection (normalized to time 0). (B) Ex vivo supplementation with IL-6 increased the MM cell populations for 3 of 5 samples tested. (C) The viability of MM cells decays ex vivo in the first 24 hours and then plateaus at 48 hours (dotted line) (n = 3). (D) Representative experiment with an MM patient sample treated with anti-myeloma drugs for 48 hours, followed by flow cytometry. Live cells are first gated by live/dead stain, followed by measuring the surviving MM cells, which are typically CD45–/CD19–/CD38+/CD138+ (CD19/CD45 gating not shown). Anti-myeloma drug treatment specifically reduces the number of MM cells at 48 hours. (E) Dose response for anti-myeloma drugs using this approach to measure drug sensitivity in cells from patients with MM. Data represent means ± SD, comparisons by 2-tailed Student t test. *P < .05; **P < .01; ****P < .0001. Bor, bortezomib; HTB, hematology tissue bank; Norm, normalized; Pom, pomalidomide; untd, untreated.$

Optimizing flow cytometry–based measurements of drug sensitivity in MM patient samples ex vivo. (A) MM cell viability after 48 hours from 4 primary samples that were thawed and separated into fractions and cultured either without selection (MNCs) or with CD138 selection (normalized to time 0). (B) Ex vivo supplementation with IL-6 increased the MM cell populations for 3 of 5 samples tested. (C) The viability of MM cells decays ex vivo in the first 24 hours and then plateaus at 48 hours (dotted line) (n = 3). (D) Representative experiment with an MM patient sample treated with anti-myeloma drugs for 48 hours, followed by flow cytometry. Live cells are first gated by live/dead stain, followed by measuring the surviving MM cells, which are typically CD45^–/CD19^–/CD38⁺/CD138⁺ (CD19/CD45 gating not shown). Anti-myeloma drug treatment specifically reduces the number of MM cells at 48 hours. (E) Dose response for anti-myeloma drugs using this approach to measure drug sensitivity in cells from patients with MM. Data represent means ± SD, comparisons by 2-tailed Student t test. *P < .05; **P < .01; ****P < .0001. Bor, bortezomib; HTB, hematology tissue bank; Norm, normalized; Pom, pomalidomide; untd, untreated.

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