PSGro-Free is a growth medium designed for humaninduced pluripotent stem cells ( iPSCs ) and humanembryonic stem cells (ESCs). It was developed atStemRD, a premier producer of growth factors, incollaboration with iPS experts at System Biosciences.
歡迎您致電 重慶市華雅干細胞技術有限公司 華雅干細胞-:
提供產品:華雅干細胞致力于為中國干細胞領域的干細胞科學研究、臍帶血/造血干細胞/臍帶/脂肪/胎盤干細胞庫、干細胞臨床轉化醫學、干細胞美容及干細胞抗衰老、免疫細胞治療、T細胞治療、臨床干細胞治療等再生醫學和個體化治療產業鏈的相關客戶提供:MSC間充質干細胞培養基、iPS/ES胚胎干細胞培養基、血清替代物、羊水培養基、基底膜基質膠細胞支架、干細胞生長分化調控因子、BioMarker 流式檢測抗體、支原體檢測試劑盒、內毒素檢測試劑盒、干細胞存儲細胞凍存袋、細胞培養瓶、USP級DMSO二甲基亞砜細胞凍存液、細胞培養相關試劑耗材儀器等種類豐富的產品,以“Focus on improving the quality of Stem Cells專注干細胞研究,提升干細胞品質”為己任,我們與客戶攜手為中國干細胞產業的發展而前行。
StemRD PGroF-500 PSGro-case, Human iPSC/ESC growth medium 人iPSC/ESC胚胎干細胞無血清培養基 產品英文描述如下:
PSGro-case is designed for human induced pluripotent stem cells (iPSCs) and human embryonic stem cells (ESCs). It is serum-free and chemically-defined as all of its ingredients are synthetic chemicals or purified recombinant proteins. PSGro-case supports feeder-free growth and maintenance of human iPSCs or human ESCs on extra-cellular matrix proteins (e.g., BD Matrigel?). Its performance is comparable to or better than the market-leading mTeSR?1 or E8 in growth rate, characteristic morphology, pluoripotent markers, normal karyotype, and differentiation to different germ layers. The robustness and stability of PSGro-case enable seeding cells over the weekend without changing medium, so you have the weekend Free.
StemRD PGroF-500 PSGro-case, Human iPSC/ESC Growth Medium 產品組分:
product
Catalog No
size
price
PSGro kit 500ml
PGro-500
500ml
¥2587
PSGro case of 12 kits
PGro-case
12X500ml
¥25987
歡迎您致電 重慶市華雅干細胞技術有限公司 華雅干細胞-:
提供產品:華雅干細胞致力于為中國干細胞領域的干細胞科學研究、臍帶血/造血干細胞/臍帶/脂肪/胎盤干細胞庫、干細胞臨床轉化醫學、干細胞美容及干細胞抗衰老、免疫細胞治療、T細胞治療、臨床干細胞治療等再生醫學和個體化治療產業鏈的相關客戶提供:MSC間充質干細胞培養基、iPS/ES胚胎干細胞培養基、血清替代物、羊水培養基、基底膜基質膠細胞支架、干細胞生長分化調控因子、BioMarker 流式檢測抗體、支原體檢測試劑盒、內毒素檢測試劑盒、干細胞存儲細胞凍存袋、細胞培養瓶、USP級DMSO二甲基亞砜細胞凍存液、細胞培養相關試劑耗材儀器等種類豐富的產品,以“Focus on improving the quality of Stem Cells專注干細胞研究,提升干細胞品質”為己任,我們與客戶攜手為中國干細胞產業的發展而前行。
Content and Storage: Components Size Storage Shelf Life
Basal Medium (on gel ice) 450mL 2 to 8°C. Keep from light 6 months
10x Supplement (dry ice) 50mL -20°C or below. Keep from light 6 months
Additional key reagents required or suggested
Extracellular matrix proteins, such as Matrigel (BD, cat# 354277) or equivalent
Accutase (Millipore, SCR005) or Collagenase IV (Invitrogen, 17104) or equivalent
ROCK inhibitor Y-27632 (StemRD, Y-01/-05/-25) or Thiazovivin (StemRD, Thia-01/-05/-25)
Medium Preparation
1. Thaw PSGro-caseSupplement overnight at 4°C. If desired, thawed Supplement can be aliquoted and stored at -20°C or below, but further freeze-thaw should be avoided.
2. To make PSGro-casecomplete medium, add 1 part of Supplement to 9 parts of Basal Medium.
3. Antibiotics can be added to the complete medium if so desired.
4. Once made, PSGro-case complete medium is stable for 2 weeks when stored in dark at 2 to 8°C.
歡迎您致電 重慶市華雅干細胞技術有限公司 華雅干細胞-:
提供產品:華雅干細胞致力于為中國干細胞領域的干細胞科學研究、臍帶血/造血干細胞/臍帶/脂肪/胎盤干細胞庫、干細胞臨床轉化醫學、干細胞美容及干細胞抗衰老、免疫細胞治療、T細胞治療、臨床干細胞治療等再生醫學和個體化治療產業鏈的相關客戶提供:MSC間充質干細胞培養基、iPS/ES胚胎干細胞培養基、血清替代物、羊水培養基、基底膜基質膠細胞支架、干細胞生長分化調控因子、BioMarker 流式檢測抗體、支原體檢測試劑盒、內毒素檢測試劑盒、干細胞存儲細胞凍存袋、細胞培養瓶、USP級DMSO二甲基亞砜細胞凍存液、細胞培養相關試劑耗材儀器等種類豐富的產品,以“Focus on improving the quality of Stem Cells專注干細胞研究,提升干細胞品質”為己任,我們與客戶攜手為中國干細胞產業的發展而前行。
StemRD PGroF-500 PSGro-case, Human iPSC/ESC growth medium 人iPSC/ESC胚胎干細胞無血清培養基 評價或評論
Coating plates with extracellular matrix proteins, such as Matrigel
Refer to manufacturers’ instructions. For BD Matrigel?, the following procedure can be followed:
1. Thaw Matrigel on ice. Dilute Matrigel 1:50 with pre‐chilled DMEM medium.
2. Immediay use the diluted Matrigel? solution to coat tissue culture plates. For a 6‐well plate, use 1 mL of diluted Matrigel? solution per well, and swirl the plate to spread the solution evenly.
3. Let the plate stand for 1 h at 37°C or overnight at 4°C.
Adaptation of growing feeder-free cells to PSGro-case
Human iPSC or ESCs that have been grown in most commonly-used hiPSC/ESC media can be easily adapted into PSGro-case. One round of 1:1 dilution of serum-containing or serum-free medium with PSGro-case is generally sufficient. However, for certain media and cell lines, a longer and more graduate adaptation may be required.
Adaptation of growing feeder-dependent cells to PSGro-case
1. Dissociate cells with Accutase, Collagenase IV or equivalent.
2. Add the previously-used medium and gently scrape off cells as aggregates. The ideal aggregate size is similar to other media and single cells from excessive pipetting may lower plating efficiency.
3. Transfer the suspension with cell aggregates into a 15 ml tube.
4. Centrifuge at 200 x g for 5 minutes at room temperature.
5. Discard Matrigel? solution, immediay add PSGro-case(e.g., 2 mL/well for a 6-well plate).
6. After centrifugation, discard the supernatant, gently resuspend the pellet with 2 mL of the previously-used medium. Note: Adding ROCK inhibitor (Y-27632 or Thiazovivin) to the medium is shown to increase the survival and plating efficiency.
7. Transfer the cell aggregates in the previously-used medium to the wells with PSGro-case. Mix. Gently.
8. Culture the cells at 37°C, with 5% CO2 and 95% humidity.
9. If cells are seeded on Friday, change medium on Monday to 4 mL PSGro-case (no need to change over the weekend). If cells are seeded on other days during the week, change next day.
1. Quickly thaw hiPSCs or hESCs in a 37°C water bath, transfer the contents to a 15 mL tube. Add 10 mL of the medium previously-used for the cells drop-wise to the tube with gentle mixing.
2. Centrifuge cells at 200 x g for 5 minutes at room temperature.
3. Discard Matrigelsolution, immediay add PSGro-case (e.g., 2 mL/well for a 6-well plate).
4. After centrifugation, discard the supernatant, gently resuspend the pellet with 2 mL of the previously-used medium. Note: Adding ROCK inhibitor (Y-27632 or Thiazovivin) to the medium is shown to increase the survival and plating efficiency.
5. Transfer the cell aggregates in the previously-used medium to the plate with PSGro-case. Mix gently.
6. Culture the cells at 37°C, with 5% CO2 and 95% humidity.
Passaging cells in PSGro-case under feeder-free condition
1. Look under microscope to identify regions of differentiation. Mark the differentiated colonies using lens marker on the bottom of the plate (e.g., 6-well plate)
2. Remove regions of differentiation by scraping with a pipette tip or by aspiration.
3. Aspirate medium from the cell culture and rinse with PBS.
4. Add 0.5 mL per well of Accutase or Collagenase IV. Let stand at room temp for 1 – 2 min.
5. Remove Accutase, and gently rinse 2 – 3 times with 2 mL PSGro-caseto remove remaining
enzymes.
6. Add 2 mL/well PSGro-case and scrape colonies off with a cell scraper.
7. Transfer the detached cell aggregates to a 15 mL conical tube and rinse the well with an additional
2 mL PSGro-case to collect any remaining aggregates.
8. Centrifuge at 200 x g for 5 minutes at room temperature.
9. Aspirate the supernatant. Resuspend pellet in 4 mL PSGro-case by pipetting gently.
Note: Adding ROCK inhibitor (Y-27632 or Thiazovivin) to the medium is shown to increase the survival and plating efficiency.
10. Plate the cell aggregates with PSGro-case onto a new plate coated with Matrigel.
11. Culture the cells at 37°C, with 5% CO2 and 95% humidity. Change medium daily.
12. If cells are seeded on Friday, change medium on Monday to 4 mL PSGro-case (no need to change over the weekend). If cells are seeded on other days during the week, change next day.
Note: If colonies are at an optimal density, cells can be split every 5 ‐ 7 days using 1:5 to 1:10 splits.
Cryopreservation of cells in PSGro-case
1. Prepare freezing medium which contains 90% PSGro-case and 10% DMSO.
Note: Adding ROCK inhibitor (Y-27632 or Thiazovivin) to the medium is shown to increase the survival and plating efficiency.
2. Perform steps 1 - 8 from Passaging cells in PSGro-case under feeder-free condition
3. Gently aspirate the supernatant taking care to keep the cell pellet intact.
4. Gently resuspend the pellet in freezing medium, taking care to leave the cell aggregates larger than would normally be done for passaging.
5. Transfer 1 mL of cell aggregates in freezing medium into each labeled cryovial.
6. Place vials into an isopropanol freezing container and place the container at ‐80°C overnight.
7. Transfer to a liquid nitrogen tank the next day for long-term storage.