StemRD  PSGro 人iPS细胞和胚胎干细胞无血清培养基

产品英文名称：StemRD PGroF-500 PSGro-case, Human iPSC/ESC Growth Medium

产品中文名称：StemRD人iPS细胞和胚胎干细胞无血清培养基,无异源级别

StemRD PGroF-500 PSGro-case, Human iPSC/ESC growth medium 人iPSC/ESC胚胎干细胞无血清培养基 500mL/套

StemRD PGroF-500 PSGro-case, Human iPSC/ESC growth medium 人iPSC/ESC胚胎干细胞无血清培养基产品特点：

★ Serum-Free 无血清成分；

★ Xeno-Free 无异种蛋白；

★ Chemically-Defined 化学成分明确；

★ Feeder-Independent 无需滋养层细胞；

★ Supports human iPSCs/ESCs 用于人iPS细胞和胚胎干细胞生长；

★ Performs Comparably to mTeSR 1 效能与mTeSR 1培养基同等。

Characteristic Morphology Human ESCs(A,B); iPSCs(C,D) in PSGro-Free®

PSGro-Free 是stemRD开发的一款针对人类胚胎干细胞（ESCs）和多能诱导干细胞（iPSCs）的生长维护胚胎干细胞无血清培养基。我们生产质量可靠的生长因子，并拥有来自SystemBiosciences的优秀iPS研究专家。PSGro-Free的所有添加组分都为化学合成或已纯化的重组蛋白，并且决不含有任何非人源的异种成分。

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.

欢迎您致电 重庆市华雅干细胞技术有限公司        华雅干细胞-：

提供产品：华雅干细胞致力于为中国干细胞领域的干细胞科学研究、脐带血/造血干细胞/脐带/脂肪/胎盘干细胞库、干细胞临床转化医学、干细胞美容及干细胞抗衰老、免疫细胞治疗、T细胞治疗、临床干细胞治疗等再生医学和个体化治疗产业链的相关客户提供：MSC间充质干细胞培养基、iPS/ES胚胎干细胞培养基、血清替代物、羊水培养基、基底膜基质胶细胞支架、干细胞生长分化调控因子、BioMarker 流式检测抗体、支原体检测试剂盒、内毒素检测试剂盒、干细胞存储细胞冻存袋、细胞培养瓶、USP级DMSO二甲基亚砜细胞冻存液、细胞培养相关试剂耗材仪器等种类丰富的产品，以“Focus on improving the quality of Stem Cells专注干细胞研究，提升干细胞品质”为己任，我们与客户携手为中国干细胞产业的发展而前行。

StemRD PGroF-500 PSGro-case, Human iPSC/ESC growth medium 人iPSC/ESC胚胎干细胞无血清培养基 产品英文描述如下：

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 产品组分：

欢迎您致电 重庆市华雅干细胞技术有限公司        华雅干细胞-：

提供产品：华雅干细胞致力于为中国干细胞领域的干细胞科学研究、脐带血/造血干细胞/脐带/脂肪/胎盘干细胞库、干细胞临床转化医学、干细胞美容及干细胞抗衰老、免疫细胞治疗、T细胞治疗、临床干细胞治疗等再生医学和个体化治疗产业链的相关客户提供：MSC间充质干细胞培养基、iPS/ES胚胎干细胞培养基、血清替代物、羊水培养基、基底膜基质胶细胞支架、干细胞生长分化调控因子、BioMarker 流式检测抗体、支原体检测试剂盒、内毒素检测试剂盒、干细胞存储细胞冻存袋、细胞培养瓶、USP级DMSO二甲基亚砜细胞冻存液、细胞培养相关试剂耗材仪器等种类丰富的产品，以“Focus on improving the quality of Stem Cells专注干细胞研究，提升干细胞品质”为己任，我们与客户携手为中国干细胞产业的发展而前行。

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-case Supplement 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-case complete 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.

欢迎您致电 重庆市华雅干细胞技术有限公司        华雅干细胞-：

提供产品：华雅干细胞致力于为中国干细胞领域的干细胞科学研究、脐带血/造血干细胞/脐带/脂肪/胎盘干细胞库、干细胞临床转化医学、干细胞美容及干细胞抗衰老、免疫细胞治疗、T细胞治疗、临床干细胞治疗等再生医学和个体化治疗产业链的相关客户提供：MSC间充质干细胞培养基、iPS/ES胚胎干细胞培养基、血清替代物、羊水培养基、基底膜基质胶细胞支架、干细胞生长分化调控因子、BioMarker 流式检测抗体、支原体检测试剂盒、内毒素检测试剂盒、干细胞存储细胞冻存袋、细胞培养瓶、USP级DMSO二甲基亚砜细胞冻存液、细胞培养相关试剂耗材仪器等种类丰富的产品，以“Focus on improving the quality of Stem Cells专注干细胞研究，提升干细胞品质”为己任，我们与客户携手为中国干细胞产业的发展而前行。

StemRD PGroF-500 PSGro-case, Human iPSC/ESC growth medium 人iPSC/ESC胚胎干细胞无血清培养基 评价或评论

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.

重庆市华雅干细胞技术有限公司主营：干细胞、细胞冻存、细胞治疗、试剂耗材 相关板块的产品，以“传递科学价值，服务科学研究”为宗旨，经过近五年的努力推广，重庆市华雅干细胞技术有限公司在全中国代理的BD科研产品、OriGen干细胞冻存袋等产品已经成功销往：上海市、北京市、重庆市、天津市、广东省的广州市、深圳市、珠海市、江西省的南昌市、四川省的成都市、绵阳、浙江省的杭州市、宁波、温州、江苏省的南京市、苏州、无锡、泰州、湖南省的长沙、陕西省的西安市、杨凌、甘肃省的兰州、宁夏的银川、新疆的乌鲁木齐、广西的南宁市、山东省的济南、青岛、东北三省 辽宁省的沈阳市、大连市、吉林省的长春市、黑龙江省的哈尔滨市等中国绝大部分省市和自治区。

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Recovery of frozen cells in PSGro-case

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 Matrigel solution, 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.

7. Change medium to 4 mL PSGro-case next day.

重庆市华雅干细胞技术有限公司主营：干细胞、细胞冻存、细胞治疗、试剂耗材 相关板块的产品，以“传递科学价值，服务科学研究”为宗旨，经过近五年的努力推广，重庆市华雅干细胞技术有限公司在全中国代理的BD科研产品、OriGen干细胞冻存袋等产品已经成功销往：上海市、北京市、重庆市、天津市、广东省的广州市、深圳市、珠海市、江西省的南昌市、四川省的成都市、绵阳、浙江省的杭州市、宁波、温州、江苏省的南京市、苏州、无锡、泰州、湖南省的长沙、陕西省的西安市、杨凌、甘肃省的兰州、宁夏的银川、新疆的乌鲁木齐、广西的南宁市、山东省的济南、青岛、东北三省 辽宁省的沈阳市、大连市、吉林省的长春市、黑龙江省的哈尔滨市等中国绝大部分省市和自治区。

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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-case to 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.

重庆市华雅干细胞技术有限公司主营：干细胞、细胞冻存、细胞治疗、试剂耗材 相关板块的产品，以“传递科学价值，服务科学研究”为宗旨，经过近五年的努力推广，重庆市华雅干细胞技术有限公司在全中国代理的BD科研产品、OriGen干细胞冻存袋等产品已经成功销往：上海市、北京市、重庆市、天津市、广东省的广州市、深圳市、珠海市、江西省的南昌市、四川省的成都市、绵阳、浙江省的杭州市、宁波、温州、江苏省的南京市、苏州、无锡、泰州、湖南省的长沙、陕西省的西安市、杨凌、甘肃省的兰州、宁夏的银川、新疆的乌鲁木齐、广西的南宁市、山东省的济南、青岛、东北三省 辽宁省的沈阳市、大连市、吉林省的长春市、黑龙江省的哈尔滨市等中国绝大部分省市和自治区。

欢迎您致电 重庆市华雅干细胞技术有限公司     华雅干细胞-：