The combined influence of substrate elasticity and surface-grafted molecules on the ex vivo expansion of hematopoietic stem and progenitor cells

Abstract

Umbilical cord blood (UCB) is an attractive source of hematopoietic stem and progenitor cells (HSPCs) for transplantation. However, the low number of HSPCs from a single UCB donor limits the direct transplantation of UCB to patients. Because little is known about the effects of the physical microenvironment on HSPC expansion, we investigated the ex vivo expansion of HSPCs cultured on biomaterials with different elasticities and grafted with different nanosegments. Polyvinylalcohol-co-itaconic acid (PVA-IA)-coated dishes with different stiffnesses ranging from a 3.7 kPa to 30.4 kPa storage modulus were used. Fibronectin or an oligopeptide (CS1, EILDVPST) was grafted onto the PVA-IA substrates. High ex vivo fold expansion of HSPCs was observed in the PVA-IA dishes grafted with fibronectin or CS1, which displayed an intermediate stiffness ranging from 12.2 kPa to 30.4 kPa. The fold expansion was more than 1.4 times higher than that cultured in tissue culture polystyrene dishes (TCPS, 12 GPa). Furthermore, HSPCs cultured in fibronectin or CS1-grafted PVA-IA-coated dishes with a stiffness of 12.2-30.4 kPa generated more pluripotent colony-forming units (CFU-GM and CFU-GEMM) than those in TCPS dishes. This result indicates that both the physical and biological properties of biomaterials affect the ex vivo expansion of HSPCs.

Substrate elasticity induces quiescence and promotes neurogenesis of primary neural stem cells—A biophysical in vitro model of the physiological cerebral milieu

“…Interestingly, these molecules are not only known for the mechanical forces they transmit but also for their various biological effects on cells in the brain (Klein et al, ; Seebeck et al, ; Williams, Furness, Walsh, & Doherty, ). Overall, elasticity constitutes an important mechanical factor influencing various cell types in the body, including embryonal stem cells (Chowdhury et al, ; Evans et al, ), mesenchymal stem cells (Engler, Sen, Sweeney, & Discher, ; Schellenberg et al, ), hematopoietic stem cells (Kumar et al, ), or cardiomyocytes (Hersch et al, ).…”

Xeno-free culture of human pluripotent stem cells on oligopeptide-grafted hydrogels with various molecular designs

“…PVA films were prepared from a previously reported method44. PVA hydrogels were prepared by crosslinking PVA films with an aqueous crosslinking solution composed of 1.0% (w/v) H 2 SO 4 , 20.0% (w/v) Na 2 SO 4 , and 1.0% (w/v) glutaraldehyde (Fig.…”

Pluripotency maintenance of amniotic fluid-derived stem cells cultured on biomaterials

“…24 A 3 mL aliquot of the PVA-IA solution was then added to a 35 mm TCPS dish and dried for a week on a clean bench to produce a film. 24 A 3 mL aliquot of the PVA-IA solution was then added to a 35 mm TCPS dish and dried for a week on a clean bench to produce a film.…”

“…24 Subsequently, the PVA hydrogels were washed with phosphate-buffered saline (PBS; pH 7.2) and immersed in a PBS solution containing 50 mg mL À1 of ECM protein (COL, FN or rVN) or ECM-derived oligopeptide (COLA, COLB, oligoFN or oligoVN) for 24 h at 4 1C (Fig. 24 Subsequently, the PVA hydrogels were washed with phosphate-buffered saline (PBS; pH 7.2) and immersed in a PBS solution containing 50 mg mL À1 of ECM protein (COL, FN or rVN) or ECM-derived oligopeptide (COLA, COLB, oligoFN or oligoVN) for 24 h at 4 1C (Fig.…”

“…Recently, in addition to biological cues such as growth factors, physical cues such as biomaterials have been recognized to affect the stem cell fate of differentiation. 24 Chowdhury et al reported that the pluripotency of mouse ESCs (mESCs) could be maintained by culture on soft hydrogels coated with collagen I (0.6 kPa) matching the intrinsic stiffness of mESCs. 19,[21][22][23] Higuchi et al investigated the expansion of hematopoietic stem cells (HSCs) on biomaterials grafted with fibronectinderived oligopeptides possessing varying degrees of stiffness.…”