CytoSelect™ 96-Well Hematopoietic Colony Forming Cell Assay (96 assays)

Product Description

• Quantify hematopoietic stem cell colony formation after just 7-10 days using a fluorescence plate reader
• No manual cell counting required
• Methylcellulose medium supports growth of hematopoietic cell colonies
• Cells can be recovered for further analysis

Hematopoietic stem cells (HSC) are well-characterized, tissue-specific stem cells that exhibit remarkable self-renewal capacity and are responsible for the life-long maintenance of the hematopoietic system. HSC are rare cells that reside in adult bone marrow where hematopoiesis is continuously taking place, but can also be found in cord blood, fetal liver, adult spleen and peripheral blood. Throughout the life span, hematopoiesis continuously replenishes the various lymphoid, myeloid and erythroid-megakaryocyte lineages, but also to maintain a small pool of HSC with the selfrenewal capacity that is capable of carrying on hematopoiesis. From HSC to mature blood cells, extensive proliferation and expansion occurs that results in the production of millions of blood cells.

When cultured in a suitable semi-solid matrix, such as methylcellulose supplemented with nutrients and cytokines, HSC or hematopoietic progenitors called colony-forming cells (CFCs) proliferate to form discrete cell clusters or colonies. Colony types include colony-forming unit-erythroid (CFU-E), burst-forming unit-erythroid (BFU-E), CFU-granulocyte, macrophage (CFU-GM) and CFUgranulocyte, erythrocyte, macrophage, megakaryocyte (CFU-GEMM).

In contrary to classical assays, Cell Biolabs CytoSelect™ 96-well Hematopoietic Colony Forming Cell Assay does not involve subjective manual counting of colonies or require a 2-3 week incubation period. Instead cells are incubated only 7-10 days in a semisolid methycellulose media before being solubilized, lysed and detected by the patented CyQuant® GR Dye in a fluorescence plate reader (see Assay Principle below).

Alternatively, viable CFCs can be easily recovered for further culturing and testing. This format provides a quantitative, high-throughput method to accurately measure HSC or hematopoietic progenitor clonogenic capability.

Assay Principle

Sample Data

Colony Formation. Human bone marrow derived CD34+ Hematopoietic Progenitor Cells were seeded at 3000 cells/well and cultured for 7 days in the presence or absence of growth factors/cytokines (hIL-3, hIL-6, hG-CSF, hEPO). Colony quantitation was determined according to the assay protocol.

Photographs were taken after 7-day culture for Panel A (without growth factor/cytokine supplement) and Panel B (with growth factor/cytokine supplement). Panel (C) demonstrates growth after 10 days with growth factors/cytokines (hemoglobin clearly visible).