Human Bone Marrow CD34+ Stem Cells
CD34 is a glycosylated transmembrane protein and represents a well-known marker for primitive blood- and bone marrow-derived progenitor cells, especially for hematopoietic and endothelial stem cells. CD34+ stem cells are multipotent and can differentiate to all hematopoietic cell types in blood. Additionally, CD34+ cells can also give rise to all lymphohematopoietic lineages even though they comprise only a small percentage of the cell population (compromising only ~1.5% of bone marrow mononuclear cell population).
Human Bone Marrow CD34+ stem cells (BM CD34+) are isolated from healthy donor's bone marrow mononuclear cells by positive selection using CD34 immunomagnetic beads.
All bone marrow is collected in heparin from fully consented IRB approved donors. All donors are tested negative of HIV, HBV and HCV.
Figure 1. Phase contrast microscopy image shows good morphology of isolated CD34+ cells. Flow cytometric analysis showed that >90% of the cells are CD34+.
CD34 is a cluster of differentiation (CD) first described independently by Civin et al. and Tindle et al. in a cell surface glycoprotein and functions as a cell-cell adhesion factor. It may also mediate the attachment of stem cells to bone marrow extracellular matrix or directly to stromal cells.
Function of CD34
The CD34 protein is a member of a family of single-pass transmembrane sialomucin proteins that show expression on early hematopoietic and vascular-associated tissue. However, little is known about its exact function.
CD34 is also an important adhesion molecule and is required for T cells to enter lymph nodes. It is expressed on lymph node endothelium, whereas the L-selectin to which it binds is on the T cell. Conversely, under other circumstances CD34 has been shown to act as molecular "Teflon" and block mast cell, eosinophil and dendritic cell precursor adhesion, and to facilitate opening of vascular lumina. Finally, recent data suggest CD34 may also play a more selective role in chemokine-dependent migration of eosinophils and dendritic cell precursors. Regardless of its mode of action, under all circumstances CD34, and its relatives podocalyxin and endoglycan, facilitates cell migration.
CD34 Tissue Distribution
Cells expressing CD34 (CD34+ cell) are normally found in the umbilical cord and bone marrow as hematopoietic cells, or in mesenchymal stem cells, endothelial progenitor cells, endothelial cells of blood vessels but not lymphatics (except pleural lymphatics), mast cells, a sub-population dendritic cells (which are factor XIIIa-negative) in the interstitium and around the adnexa of dermis of skin, as well as cells in soft tissue tumors like dermatofibrosarcoma protuberans (DFSP), gastrointestinal stromal tumors (GISTs), solitary fibrous tumor (SFT), hemangiopericytoma (HPC), and to some degree in malignant peripheral nerve sheath tumors (MPNSTs), etc. The presence of CD34 on non-hematopoietic cells in various tissues has been linked to progenitor and adult stem cell phenotypes.
It is important to mention that Long-Term Hematopoietic Stem Cells (LT-HSCs) in mice and humans are the hematopoietic cells with the greatest self-renewal capacity. Human HSCs express the CD34 marker.
CD34 is expressed in roughly 20% of murine hematopoietic stem cells, and can be stimulated and reversed.
CD34+ cells may be isolated from blood samples using immunomagnetic or immunofluorescent methods.
Antibodies are used to quantify and purify hematopoietic progenitor stem cells for research and for clinical bone marrow transplantation. However, counting CD34+ mononuclear cells may overestimate myeloid blasts in bone marrow smears due to B lymphocyte precursors and CD34+ megakaryocytes.
Cells observed as CD34+ and CD38- are of an undifferentiated, primitive form; i.e., they are multipotential hemopoietic stem cells. Thus, because of their CD34+ expression, such undifferentiated cells can be sorted out.
In tumors, CD34 is found in alveolar soft part sarcoma, preB-ALL (positive in 75%), AML (40%), AML-M7 (most), dermatofibrosarcoma protuberans, gastrointestinal stromal tumors, giant cell fibroblastoma, granulocytic sarcoma, Kaposi’s sarcoma, liposarcoma, malignant fibrous histiocytoma, malignant peripheral nerve sheath tumors, meningeal hemangiopericytomas, meningiomas, neurofibromas, schwannomas, and papillary thyroid carcinoma.
A negative CD34 may exclude Ewing's sarcoma/PNET, myofibrosarcoma of the breast, and inflammatory myofibroblastic tumors of the stomach.
Injection of CD34+ hematopoietic Stem Cells has been clinically applied to treat various diseases including Spinal Cord Injury, Liver Cirrhosis and Peripheral Vascular disease. Research has shown that CD34+ cells are relatively more in men than in women in the reproductive age among Spinal Cord Injury victims.
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