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Nutrifreez d10

Manufactured by Sartorius
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The NutriFreez D10 is a laboratory equipment designed for freeze-drying samples. It provides a controlled environment for the removal of water from various materials through the process of sublimation.

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Lab products found in correlation

Fibroblast growth factor (fgf), by Thermo Fisher Scientific (2 mentions) Tryple, by Thermo Fisher Scientific (2 mentions) Stemdiff mesenchymal progenitor kit, by STEMCELL (2 mentions) Penicillin streptomycin p s, by Thermo Fisher Scientific (2 mentions) Fetal bovine serum (fbs), by Corning (2 mentions) Tryple select enzyme 10x, by Thermo Fisher Scientific (1 mentions) Cytotune 2, by Thermo Fisher Scientific (1 mentions) Mtesr1, by STEMCELL (1 mentions)

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NutriFreez® D10 Cryopreservation Medium Nutrifreez

4 protocols using nutrifreez d10

1

hiPSC Differentiation into MSCs

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hiPSCs were differentiated into MSCs following manufacturer’s protocol in the STEMdiff Mesenchymal Progenitor Kit (STEMCELL Technologies). Briefly, hiPSCs were passaged under standard conditions 2 days prior to starting a 4-day induction into early mesodermal progenitors. To derive mesenchymal progenitors, cells were then switched to the supplied MesenCult medium and passaged without using animal components to reach an approximately 70% confluency each time.
After the 21-day differentiation period, hiPSC-derived MSCs (hiMSCs) were cultured in MSC medium comprised of hgDMEM, 10% (v/v) fetal bovine serum (Corning), 1% (v/v) penicillin/streptomycin (P/S; Gibco), and 0.1 ng/mL fibroblast growth factor (Peprotech) with media changes twice per week on uncoated tissue culture plastic. hiMSCs were dissociated using TrypLE (Gibco) and cryopreserved with NutriFreez D10 (Biological Industries).
Wu J.Y., Yeager K., Tavakol D.N., Morsink M., Wang B., Soni R.K., Hung C.T, & Vunjak-Novakovic G. (2023). Directed differentiation of human iPSCs into mesenchymal lineages by optogenetic control of TGF-β signaling. Cell reports, 42(5), 112509.
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2

Isolation and Cryopreservation of Amniotic Epithelial Cells

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This study was approved by the Ethics Committee of the Graduate School of Medicine, Tohoku University (Permission number: 2019‐1‐430). The amniotic membrane was collected with the consent of a pregnant woman undergoing a scheduled cesarean section at the Department of Obstetrics, Tohoku University Hospital. The amniotic membrane was collected from the placenta, which was removed after delivery by cesarean section. The isolation of AE cells was performed according to the protocol of Gramignoli et al.17 TrypLE Select Enzyme (10X) (Thermo Fisher Scientific, Waltham, MA, USA) was added to 1 g of amniotic membrane and incubated at 35 rpm for 30 min at 37°C to isolate AE cells. The AE cells that had undergone the filtration process were placed in the cell preservation solution NutriFreez D10 (Biological Industries, Cromwell, CT, USA) and stored frozen at –80°C or lower.
Cryopreserved AE cells were thawed at 37°C for 1 min and washed with Dulbecco's phosphate‐buffered saline (PBS). Two milliliters of normal saline were then added per 1.0 × 10⁷ of AE cells.
Konno Y., Sato C., Tokodai K., Saito M., Hoshiai T., Taniyama Y., Okamoto H., Fukutomi T., Ozawa Y., Ujiie N., Koseki K., Ando R., Takahashi K., Shinozaki Y., Unno M, & Kamei T. (2022). A potential preventive method for scar stenosis after esophageal endoscopic mucosal resection using human amniotic epithelial cells in a porcine model. DEN Open, 2(1), e104.
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3

hiPSC Differentiation into MSCs

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hiPSCs were differentiated into MSCs following manufacturer’s protocol in the STEMdiff Mesenchymal Progenitor Kit (STEMCELL Technologies). Briefly, hiPSCs were passaged under standard conditions 2 days prior to starting a 4-day induction into early mesodermal progenitors. To derive mesenchymal progenitors, cells were then switched to the supplied MesenCult medium and passaged without using animal components to reach an approximately 70% confluency each time.
After the 21-day differentiation period, hiPSC-derived MSCs (hiMSCs) were cultured in MSC medium comprised of hgDMEM, 10% (v/v) fetal bovine serum (Corning), 1% (v/v) penicillin/streptomycin (P/S; Gibco), and 0.1 ng/mL fibroblast growth factor (Peprotech) with media changes twice per week on uncoated tissue culture plastic. hiMSCs were dissociated using TrypLE (Gibco) and cryopreserved with NutriFreez D10 (Biological Industries).
Wu J.Y., Yeager K., Tavakol D.N., Morsink M., Wang B., Soni R.K., Hung C.T, & Vunjak-Novakovic G. (2023). Directed differentiation of human iPSCs into mesenchymal lineages by optogenetic control of TGF-β signaling. Cell reports, 42(5), 112509.
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4

Generation and Characterization of Human iPSCs

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Human induced pluripotent stem cells (hiPSC) were generated at CR-CHUSJ Stem Cell core or in-house. The hiPSC lines SJi3252C2 and SJ3013C2 were derived from human fibroblasts using Cytotune 1.0 (Invitrogen) and were previously characterized. 80 EU03.C2, and EU148.C5 were derived from human PBMC with Cytotune 2.0 (A16517, Invitrogen). The hiPSC eGFP line (SEC61BGFP/AICS-0010) was acquired from Allen Institute for Cell Science. 81 The abbreviations used for the above iPSC lines in this report are as follows: C1 (SJi3252C2), C2 (SJ3013C2), C3 (EU03.C2), C4 (AICS-0010), and C5 (EU148.C5). The hiPSCs were cultured in hypoxic condition (5%CO 2 , 5%O 2 , 37 C incubator) until passage 15-20, otherwise all cells and derivatives were cultured in normoxic condition (5% CO 2 , 37 C incubator). They were cultured with mTeSR1 (85850, StemCell Technologies) with 1X penicillin-streptomycin (450-201-EL, MultiCell) and hESC-qualifed Matrigel (354277, Corning). Matrigel was coated onto Nunc Delta surface plates (14-832-11, Thermo Scientific) as per manufacturer recommendation. The cells were passaged as small clusters using 0.5mM ethylenediaminetetraacetic acid (EDTA) in phosphate buffered saline (PBS). The cells were cryopreserved with NutriFreezD10 (05-713-1E, Biological Industries) as per manufacturer recommendation.
Song A.T., Sindeaux R.H.M., Li Y., Affia H., Agnihotri T., Leclerc S., van Vliet P.P., Colas M., Guimond J.V., Patey N., Feulner L., Joyal J.S., Haddad E., Barreiro L, & Andelfinger G. (2024). Developmental role of macrophages modeled in human pluripotent stem cell-derived intestinal tissue. Cell reports, 43(1).
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