ll

Previews
Bowie, M.B., Kent, D.G., Dykstra, B., McKnight, potential occurs at the level of hematopoietic Reprogramming that Begins before Birth. Cell
K.D., McCaffrey, L., Hoodless, P.A., and Eaves, stem cells. Cell 62, 863–874. Stem Cell 27, this issue, 732–747.
C.J. (2007). Identification of a new intrinsically
timed developmental checkpoint that reprograms Mu€ller, A.M., Medvinsky, A., Strouboulis, J.,
Kikuchi, K., and Kondo, M. (2006). Developmental
key hematopoietic stem cell properties. Proc. Grosveld, F., and Dzierzak, E. (1994).
switch of mouse hematopoietic stem cells from
Natl. Acad. Sci. USA 104, 5878–5882. fetal to adult type occurs in bone marrow after
Development of hematopoietic stem cell activity
in the mouse embryo. Immunity 1, 291–301.
birth. Proc. Natl. Acad. Sci. USA 103,
Copley, M.R., and Eaves, C.J. (2013). 17852–17857. Pietras, E.M., Reynaud, D., Kang, Y.A., Carlin, D.,
Developmental changes in hematopoietic stem Calero-Nieto, F.J., Leavitt, A.D., Stuart, J.M.,
cell properties. Exp. Mol. Med. 45, e55. Li, Y., Kong, W., Yang, W., Patel, R.M., Casey, Göttgens, B., and Passegué, E. (2015).
E.B., Okeyo-Owuor, T., White, J.M., Porter, S.N., Functionally Distinct Subsets of Lineage-Biased
Ikuta, K., Kina, T., MacNeil, I., Uchida, N., Peault, Morris, S.A., and Magee, J.A. (2020). Single-Cell Multipotent Progenitors Control Blood Production
B., Chien, Y.H., and Weissman, I.L. (1990). A devel- Analysis of Neonatal HSC Ontogeny Reveals in Normal and Regenerative Conditions. Cell
opmental switch in thymic lymphocyte maturation Gradual and Uncoordinated Transcriptional Stem Cell 17, 35–46.

Dynamic Hydrogels
for Investigating Vascularization
Gordana Vunjak-Novakovic1,*
1Columbia University, New York, NY 10032, USA

*Correspondence: [email protected]
https://doi.org/10.1016/j.stem.2020.10.009

Extracellular matrix is known to regulate vascularization by a sequence of multiple factors including mechan-
ical forces. In this issue of Cell Stem Cell, Wei et al. (2020) investigate the roles of matrix biomechanics on
early stages of vasculogenesis by using hydrogels with tunable stiffness and stress relaxation.

Cell fate and function, in vivo and in vitro, tissue culture. While vascular phenotypes field has also started to incorporate
are regulated by the entire context of the change with the developmental stage, another facet of control where materials
cellular environment, through cascades parent tissue type, and the state of health that change their properties in response
of interacting molecular, structural, and or disease, achieving angiogenesis and to cell-generated signals are being
physical signals that change in space vasculogenesis is critically important explored. The present study is an impor-
and time. The cells in turn regulate their both for the survival of implanted tissues tant step toward establishing and utilizing
environment as they secrete cytokines, and for designing tissue models for bio- dynamic cellular environments. The au-
build and degrade the extracellular ma- logical research. In this study, the focus thors explored if the activation of specific
trix, influence the surrounding cells, and is on the effects of the mechanical envi- mechanosensing mechanisms in cultured
generate physical forces. These two-way ronment, established by a hydrogel used vascular cells, in conjunction with matrix
interactions between the cells and their to encapsulate cells, on the initiation, pro- remodeling, can be utilized to induce the
environment are instrumental for native gression, and outcomes of early stages of assembly of vascular networks. To this
tissue development, phenotypic changes vasculogenesis. end, they developed a viscoelastic hydro-
associated with diseases, and the assem- It is well known that the extracellular gel with dynamic crosslinks that pro-
bly and function of engineered tissues. In matrix provides not only a three-dimen- moted cellular contractility and enabled
recent years, in vitro models of increas- sional setting for the cultured cells but formation of large focal adhesions and
ingly high biological fidelity have been also serves as a major source of regulato- robust vascular networks. This sequence
developed to enable controllable studies ry signals that determine vascular of events could not be initiated using
under conditions more closely emulating development and function, including otherwise identical hydrogels with static
the in vivo milieu (Paek et al., 2019; Ro- physical forces (Discher et al., 2009). covalent crosslinks.
naldson-Bouchard and Vunjak-Nova- With this understanding, the field has To decouple the effects of stiffness and
kovic, 2018; Skylar-Scott et al., 2019). gradually moved from generic, inert, matrix dynamics, the authors conducted
In this issue of Cell Stem Cell, the Wei and nondegradable biomaterials toward experiments with dynamic and static
et al. (2020) article reports a study of designing those with tunable properties matrices that had the same initial stiffness
vascularization, which remains one of that can be tailored to control cell differen- at two different levels: low (soft matrix)
the universal challenges in cell and tiation and assembly. In recent years, the and high (stiff matrix). Cultivation of

Cell Stem Cell 27, November 5, 2020 ª 2020 Elsevier Inc. 697
 ll
Previews

endothelial cells in dynamic hydrogels re- tion, by virtue of their elastic bonds, works over longer periods of time. Inves-
sulted in remarkable effects on the length such that the cells are relieved of external tigating the other sources of human cells,
and volume of vascular tubes, both of forces and allowed to elongate, contract, including stem cells, that give rise to
which increased 10-fold relative to the and migrate inside the hydrogel and to vascular networks would also be of inter-
vascular tubes formed in static hydrogels. form focal adhesions. In contrast, the co- est. Finally, this approach could be
In addition, vascular sprouting and valent bonds in static hydrogels maintain extended to vascularization of devel-
branching were observed only in dynamic the initial levels of stress in the hydrogel oping and regenerating tissues, as well
hydrogels. Importantly, these results were surrounding the cells. Importantly, this dif- as tumor tissues, which display a broad
not due to the differences in stiffness or ference is maintained in both the soft and range of biomechanical environments.
diffusion rates in dynamic and static stiff hydrogels, suggesting that the One could envision that the use of re-
hydrogels. observed behaviors are independent of porters designed to mark some of the
This study also showed that the inhibi- stiffness. key steps in vasculogenesis would lead
tion of cell contraction did not decrease The model proposes that the interac- to dynamic and noninvasive longitudinal
integrin intensity and number, but it did tions of integrins with the Arg-Gly-Asp studies of the events involved in vascu-
affect the integrin clustering, which is crit- (RGD)binding sites in dynamic hydrogels larization under normal and pathological
ical for the cellular protrusions and help recruit focal adhesion proteins, conditions.
sprouting that lead to the formation of including FAK. The resulting clustering
the vascular bed. Collectively, the data of integrins, mediated by actin contrac-
suggest that integrin clustering serves as tility, and the recruitment of vinculin REFERENCES
a checkpoint for vasculogenesis in an in- then lead to the formation of large and
tensity- and timing-dependent manner. stable focal adhesions. Finally, the upre- Discher, D., Mooney, D.J., and Zandstra, P.W.
(2009). Growth factors, matrices, and forces
These findings are consistent with previ- gulation of MT1-MMP, MMP-1, and
combine and control stem cells. Science 324,
ous studies that demonstrated that the MMP-9 promote matrix degradation, 1673–1677.
formation of focal adhesions in endothe- further facilitating cell sprouting and the
lial cells is initiated by activation of FAK formation of nascent vascular networks. Iruela-Arispe, M.L., and Davis, G.E. (2009). Cellular
and molecular mechanisms of vascular lumen for-
and contractility-mediated integrin clus- In support of the model, the rigidity of mation. Dev. Cell 16, 222–231.
tering (Yu et al., 2011). The hydrogel dy- covalent bonds in static hydrogels sup-
namics also enhanced the remodeling of pressed the recruitment of focal adhe- Paek, J., Park, S.E., Lu, Q., Park, K.T., Cho, M., Oh,
J.M., Kwon, K.W., Yi, Y.S., Song, J.W., Edelstein,
extracellular matrix by increasing both sion proteins and thereby inhibited cell H.I., et al. (2019). Microphysiological Engineering
the degradation of the provisional matrix contractility and the subsequent series of Self-Assembled and Perfusable Microvascular
Beds for the Production of Vascularized Three-
by Matrix metallopeptidases (MMPs) and of events leading to the formation of mi- Dimensional Human Microtissues. ACS Nano 13,
the deposition of collagen IV and laminin, crovessels. Vacuole and lumen formation 7627–7643.
the two components of the basement were also observed in static hydrogels,
Ronaldson-Bouchard, K., and Vunjak-Novakovic,
membrane. Another previous finding sup- but they were suppressed by the lack G. (2018). Organs-on-a-Chip: A Fast Track for
ported by this study is that the dynamic of stress relaxation around the cells. Engineered Human Tissues in Drug Development.
remodeling of the matrix is necessary for The delayed integrin clustering and Cell Stem Cell 22, 310–324.
the initiation and progression of vasculo- focal adhesion formation could not be
Skylar-Scott, M.A., Uzel, S.G.M., Nam, L.L.,
genesis by allowing endothelial cell (EC) compensated by prolonging the time in Ahrens, J.H., Truby, R.L., Damaraju, S., and
motility (Iruela-Arispe and Davis, 2009). culture. Lewis, J.A. (2019). Biomanufacturing of organ-spe-
cific tissues with high cellular density and
The authors integrated the experi- This study provides a conceptual embedded vascular channels. Sci. Adv. 5,
mental data and observations into a approach and also engineers an impor- eaaw2459.
mechanistic model that describes regula- tant tool: hydrogels capable of providing
Wei, Z., Schnellmann, R., Pruitt, H.C., and Gerecht,
tion of the initiation, progression, and out- stress relaxation in a broad and physio- S. (2020). Hydrogel Network Dynamics Regulate
comes of early stages of vascular network logically relevant range of stiffnesses. Vascular Morphogenesis. Cell Stem Cell 27, this
formation in the dynamic and static hy- Fast degradation of hydrogels limited issue, 798–812.
drogels used to encapsulate vascular this study to early stages of vasculogen- Yu, C.H., Law, J.B., Suryana, M., Low, H.Y., and
cells (Figure 7G in Wei et al., 2020). The esis, i.e., within the first 7 days. Future Sheetz, M.P. (2011). Early integrin binding to Arg-
fundamental difference between the two studies would benefit from hydrogels de- Gly-Asp peptide activates actin polymerization
and contractile movement that stimulates outward
types of hydrogels is that the dynamic hy- signed to allow studies of the phenotypic translocation. Proc. Natl. Acad. Sci. USA 108,
drogels have the ability for stress relaxa- stability and function of vascular net- 20585–20590.

698 Cell Stem Cell 27, November 5, 2020