Abstract
In this methodological paper, lyophilized fibroin-coated alginate microcarriers (LFAMs) proposed as mesenchymal stem cells (MSCs) delivery systems and optimal MSCs seeding conditions for cell adhesion rate and cell arrangement, was defined by a Design of Experiment (DoE) approach. Cells were co-incubated with microcarriers in a bioreactor for different time intervals and conditions: variable stirring speed, dynamic culture intermittent or continuous, and different volumes of cells-LFAMs loaded in the bioreactor. Intermittent dynamic culture resulted as the most determinant parameter; the volume of LFAMs/cells suspension and the speed used for the dynamic culture contributed as well, whereas time was a less influencing parameter. The optimized seeding conditions were: 98 min of incubation time, 12.3 RPM of speed, and 401.5 µL volume of cells-LFAMs suspension cultured with the intermittent dynamic condition. This DoE predicted protocol was then validated on both human Adipose-derived Stem Cells (hASCs) and human Bone Marrow Stem Cells (hBMSCs), revealing a good cell adhesion rate on the surface of the carriers. In conclusion, microcarriers can be used as cell delivery systems at the target site (by injection or arthroscopic technique), to maintain MSCs and their activity at the injured site for regenerative medicine.
| Original language | English |
|---|---|
| Journal | Pharmaceutics |
| Volume | 10 |
| Issue number | 4 |
| DOIs | |
| Publication status | Published - Oct 24 2018 |
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Silk/Fibroin Microcarriers for Mesenchymal Stem Cell Delivery : Optimization of Cell Seeding by the Design of Experiment. / Perucca Orfei, Carlotta; Talò, Giuseppe; Viganò, Marco; Perteghella, Sara; Lugano, Gaia; Fabro Fontana, Francesca; Ragni, Enrico; Colombini, Alessandra; De Luca, Paola; Moretti, Matteo; Torre, Maria Luisa; de Girolamo, Laura.
In: Pharmaceutics, Vol. 10, No. 4, 24.10.2018.Research output: Contribution to journal › Article
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TY - JOUR
T1 - Silk/Fibroin Microcarriers for Mesenchymal Stem Cell Delivery
T2 - Optimization of Cell Seeding by the Design of Experiment
AU - Perucca Orfei, Carlotta
AU - Talò, Giuseppe
AU - Viganò, Marco
AU - Perteghella, Sara
AU - Lugano, Gaia
AU - Fabro Fontana, Francesca
AU - Ragni, Enrico
AU - Colombini, Alessandra
AU - De Luca, Paola
AU - Moretti, Matteo
AU - Torre, Maria Luisa
AU - de Girolamo, Laura
PY - 2018/10/24
Y1 - 2018/10/24
N2 - In this methodological paper, lyophilized fibroin-coated alginate microcarriers (LFAMs) proposed as mesenchymal stem cells (MSCs) delivery systems and optimal MSCs seeding conditions for cell adhesion rate and cell arrangement, was defined by a Design of Experiment (DoE) approach. Cells were co-incubated with microcarriers in a bioreactor for different time intervals and conditions: variable stirring speed, dynamic culture intermittent or continuous, and different volumes of cells-LFAMs loaded in the bioreactor. Intermittent dynamic culture resulted as the most determinant parameter; the volume of LFAMs/cells suspension and the speed used for the dynamic culture contributed as well, whereas time was a less influencing parameter. The optimized seeding conditions were: 98 min of incubation time, 12.3 RPM of speed, and 401.5 µL volume of cells-LFAMs suspension cultured with the intermittent dynamic condition. This DoE predicted protocol was then validated on both human Adipose-derived Stem Cells (hASCs) and human Bone Marrow Stem Cells (hBMSCs), revealing a good cell adhesion rate on the surface of the carriers. In conclusion, microcarriers can be used as cell delivery systems at the target site (by injection or arthroscopic technique), to maintain MSCs and their activity at the injured site for regenerative medicine.
AB - In this methodological paper, lyophilized fibroin-coated alginate microcarriers (LFAMs) proposed as mesenchymal stem cells (MSCs) delivery systems and optimal MSCs seeding conditions for cell adhesion rate and cell arrangement, was defined by a Design of Experiment (DoE) approach. Cells were co-incubated with microcarriers in a bioreactor for different time intervals and conditions: variable stirring speed, dynamic culture intermittent or continuous, and different volumes of cells-LFAMs loaded in the bioreactor. Intermittent dynamic culture resulted as the most determinant parameter; the volume of LFAMs/cells suspension and the speed used for the dynamic culture contributed as well, whereas time was a less influencing parameter. The optimized seeding conditions were: 98 min of incubation time, 12.3 RPM of speed, and 401.5 µL volume of cells-LFAMs suspension cultured with the intermittent dynamic condition. This DoE predicted protocol was then validated on both human Adipose-derived Stem Cells (hASCs) and human Bone Marrow Stem Cells (hBMSCs), revealing a good cell adhesion rate on the surface of the carriers. In conclusion, microcarriers can be used as cell delivery systems at the target site (by injection or arthroscopic technique), to maintain MSCs and their activity at the injured site for regenerative medicine.
U2 - 10.3390/pharmaceutics10040200
DO - 10.3390/pharmaceutics10040200
M3 - Article
C2 - 30352986
VL - 10
JO - Pharmaceutics
JF - Pharmaceutics
SN - 1999-4923
IS - 4
ER -