Tracing Hematopoietic Progenitor Cell Neutrophilic Differentiation via Raman Spectroscopy.

A major challenge to experimental studies and therapeutic uses of hematopoietic stem cells (HSC) is the limited options for analytical tools that can reliably resolve functional differences in heterogeneous HSC subpopulations at the single cell level. Currently available methods require the use of external labels and/or separate clonogenic and transplantation assays to identify bona fide stem cells, necessitating the harvest of bulk cell populations and long incubation times that obscure how individual HSCs dynamically respond to exogenous and endogenous stimuli. In this study, we employ Raman spectroscopy to noninvasively resolve the dynamics of individual differentiating hematopoietic progenitor cells during the course of neutrophilic differentiation.

Identifying States along the Hematopoietic Stem Cell Differentiation Hierarchy with Single Cell Specificity via Raman Spectroscopy.

A major challenge for expanding specific types of hematopoietic cells ex vivo for the treatment of blood cell pathologies is identifying the combinations of cellular and matrix cues that direct hematopoietic stem cells (HSC) to self-renew or differentiate into cell populations ex vivo. Microscale screening platforms enable minimizing the number of rare HSCs required to screen the effects of numerous cues on HSC fate decisions. These platforms create a strong demand for label-free methods that accurately identify the fate decisions of individual hematopoietic cells at specific locations on the platform.

Secondary ion mass spectrometry and Raman spectroscopy for tissue engineering applications.

Identifying the matrix properties that permit directing stem cell fate is crucial for expanding desired cell lineages ex vivo for disease treatment. Such efforts require knowledge of matrix surface chemistry and the cell responses they elicit. Recent progress in analyzing biomaterial composition and identifying cell phenotype with two label-free chemical imaging techniques, TOF-SIMS and Raman spectroscopy are presented.

Identifying the lineages of individual cells in cocultures by multivariate analysis of Raman spectra.

The cellular and matrix cues that induce stem cell differentiation into distinct cell lineages must be identified to permit the ex vivo expansion of desired cell populations for clinical applications. Combinatorial biomaterials enable screening multiple different microenvironments while using small numbers of rare stem cells. New methods to identify the phenotypes of individual cells in cocultures with location specificity would increase the efficiency and throughput of these screening platforms.