Hematopoietic Stem Cell Culture

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Hematopoietic stem cells (HSCs) give rise to all blood cell types including myeloid and lymphoid lineages. Myeloid cells include monocytes, macrophages, neutrophils, basophils, eosinophils, erythrocytes, dendritic cells, and megakaryocytes or platelets. Lymphoid cells include T cells, B cells, and natural killer cells. HSCs are found in the bone marrow of adults, umbilical cord blood, placenta, and mobilized peripheral blood. Recent advances in hematopoietic stem cell biology have resulted in the use of HSC transplants in the treatment of cancers and other immune system disorders. We offer a broad range of tools and technologies to culture hematopoietic stem cells including Stemline® optimized serum-free expansion media, growth factors, cytokines and a broad range or HSC related antibodies and characterization kits.

Figure 1. Stages of hematopoiesis. Through the process of hematopoiesis, all cellular blood components are derived from hematopoietic stem cells.

 

 Stromal Cell Lines For T-Cell Differentiation

T lymphocytes develop in the thymus from bone-marrow derived hematopoietic stem (HSCs) and progenitor cells (HSPCs).  In vitro, various model systems have been employed to mimic thymic function supportive of T-cell development.  The classic OP9-DL1 culture system utilizes a coculture of HSPC with a murine bone marrow-derived stromal cell line (OP9) that ectopically expresses the Notch ligand, Delta-like 1 (Dll1).  The OP9-DL1 culture system utilizes fetal calf serum and thus contributes to variable efficiencies in T-cell differentiation and moreover suffers from low maturation rates. Recently, a new serum-free artificial thymic organoid (ATO) system was developed that supports highly efficient and reproducible in vitro differentiation and positive selection of human T-cells from multiple sources of HSPCs, including cord blood, bone marrow and peripheral blood CD34+ HSPCs.  A key component of the ATO system is the MS5-DLL1 cell line which eliminates the need for primary thymic tissues.  To form organoids, MS5-DLL1 stromal cells are aggregated with HSPCs by centrifugation and cultured on a Millicell® cell culture insert in defined serum-free media.

Reference: Amélie Montel-Hagen et al. Generation of mature T cells from human hematopoietic stem/progenitor cells in artificial thymic organoids. Nat Methods. 2017 May; 14(5): 521–530.

Product Description Part Number
MS5-mDLL1 Stromal Cell Line (Mouse DLL1/mStrawberry) SCC166
MS5-hDLL1 Stromal Cell Line (Human DLL1/eGFP) SCC167

 

 Hematopoietic Stem Cell Media and Cytokines

Stemline® II Hematopoietic Stem Cell Media

Developed to promote the optimal expansion of human hematopoietic stem cells (HSC) from bone marrow, mobilized peripheral blood, and cord blood, Stemline® Hematopoietic Stem Cell Expansion Medium demonstrates higher total nucleated cell (TNC) fold increases than other commercially available serum-free media formulations.

Features and Benefits

  • Serum-free formulation
  • Enhanced expansion from cord blood CD34+ cells
  • Expands cells from all appropriate hematopoietic lineages in a colony-forming unit.
  • Tested extensively in 7-day and 14-day growth assays.
  • Produced in a GMP state-of-the-art facility with an available Device Master File (DMF).

Published in over 1000 Publications

Figure 2. Stemline® demonstrates superior expansion of bone marrow hematopoietic stem cells (HSC). To test the ability of Stemline® II Hematopoietic Stem Cell Expansion Medium to expand CD34+ HSCs, in - house researchers at our lab and the University of Kentucky designed a bench-scale expansion assay. Cells were seeded into the wells of 24-well tissue culture plates. One milliliter of medium was added to each well with the appropriate cytokines to stimulate growth (100ng/mL each of TPO, SCF and G-CSF). Each condition was performed in triplicate and seeded with 10,000 cells per ml in each well. Cells were counted on day 14 and the fold increase was determined by cells final/cells initial. HSCs from bone marrow cultured in Stemline® and Stemline®II demonstrated superior expansion to those grown in other serum-free HSC media.  

Cytokines and Growth Factors

Numerous immune related cytokines are involved in the regulation of hematopoiesis within a complex network of positive and negative regulators. Some cytokines have very narrow lineage specificities of their actions, while many others have rather broad and overlapping specificity ranges. Listed within this section are the cytokines and growth factors whose action appears to be the stimulation of or regulation of hematopoietic stem cell development:

Popular Hematopoietic Stem Cell Related Cytokines

Interleukins
GM-CSF
G-CSF
M-CSF
EPO TPO SCF Flt-3 Ligand

Browse all growth factors and cytokines

HumanKine® Xeno-Free Growth Factors and Cytokines

We are proud to bring you HumanKine® growth factors and cytokines. These highly purified reagents are developed from an efficient human cell-based technology, and are excellent choices for your critical inflammation, cancer, stem cell, and antibody development research applications. 

HumanKine® Growth Factors are: 

  • Produced in HEK293 cells
  • Recombinant, animal component-free
  • Have authentic human glycosylation and  post-translational modification patterns
  • Have higher biological activity vs. E.coli derived recombinant proteins
  • Highly pure, low endotoxin proteins

 

 Hematopoietic Stem Cell Characterization

Hematopoietic Stem Cell Antibodies

Popular Hematopoietic Stem Cell Antibodies
 

CD34 CD38
CD90 CD133
CD105 CD45 c-Kit SCA-1 Stem Cell Factor ALDH

AldeRed™ ALDH Detection Assay

Hematopoietic stem cell express elevated levels of aldehyde dehydrogenase (ALDH). ALDH activity has been used as a marker for the quality of hematopoietic stem cell transplants. Historically, ALDH levels have been measured using live cell-permeable dyes that emit in the green region of the electromagnetic spectrum (512 nm). Thus, the reagent cannot be simultaneously utilized in cells or mice expressing green-fluorescent proteins emitting in the green fluorescent spectrum. The AldeRed™ ALDH Detection Assay is a red-shifted fluorescent substrate for ALDH used for labelling viable ALDH positive HSCs leaving the green emission channel available for further experimentation.

 

Read about AldeRed™ in Nature

AldeRed™ Features and Benefits:

  • Red-shifted assay leaves green channel available for further experimentation
  • Live stem cell identification enables flow sorting of rare cell populations
  • Rapid enzymatic assay protocol

 

Figure 2. AldeRed™ can label hematopoietic stem cells in live cultures. AldeRed™ 588-A is a fluorescent and non-toxic ALDH substrate that diffuses freely into live intact and viable cells, but remains trapped inside the cells once converted by ALDH into the corresponding acid.

 

Cell Culture