[New Substrate for cell culture] Recombinant Laminin 511-E8 fragments "iMatrix-511"_최적의 줄기세포(ES/iPS 세포) 배양 기질 - 코아사이언스

iMatrix-511 

Recombinant Laminin 511-E8 fragments

for cell culture of various cells - especially iPS / ES cells 

iMatrix511_NIP_140708_CoreSciences.pdf

라미닌은 동물의 기저막에 존재하는 세포외 기질의 구성물질 중 하나로 세포 부착 및 증식에 ​​깊이 관여 하는 것으로 알려져 있습니다.

본 제품은 라미닌 511 - E8 fragment와 동일한 서열을 갖는 재조합 단백질이며, 다양한 종류의 세포 접착 및 신장을 자극 할 수 있는 cell culture substrate 입니다. 오사카와 교토 대학의 공동 연구에 의해 지금까지 취급이 매우 어렵다고 여겨져 온 사람 iPS 세포와 인간 ES 세포의 배양에서도 안전하고 고효율로 배양 할 수 있는 것으로 밝혀졌습니다. 

 

Features 
■ 조작이 매우 간단!
■ 다양한 종류의 세포 배양에 사용 가능! - 특히 iPS / ES cells
■ 높은 세포 생존율 및 증식율 실현!
■ 재조합 단백질(CHO-S 세포 유래)을 사용하므로 이종 성분(xeno-components) 혼입의 위험성이 낮음!
■ 175ug의 iMatrix-511으로 약 6 well plate 2~6장 코팅 가능.

■ 인간 ES/iPS 세포의 배양에서 지금까지 어려웠던 feeder-free, 단일 세포 계대가 가능!

 

Ordering Information

Product Name	Size	Cat.#	Storage
iMatrix-511	2x175μg	892001	4℃
iMatrix-511	6x175μg	892002	4℃

 

* Activity: The dissociation constant of the binding activity with integrin α6β1 is under 10nM.

* Application: iMatrix-511 is able to use as cell culture substrate for various cell types including ES/iPS cells.

Introduction

iMatrix-511, is the recombinant human Laminin-511 E8 fragment, produced by CHO-S cells and it can reduce the risk of the xeno-components contamination in the culture. Handling is very easy - just dilute in water, coat it in the culture dish or plate and incubate at room temparature for 3 hours or at 4°C over night.

 

Laminin is one of the extra cellular matrix (ECM)s located in the basement membrane of animals, and play an important role for cell adhesion and proliferation.

 

Laminin is composed of three parts ; alpha, beta, and gamma - chain and 15 types of laminin is already known. Among them, Laminin composed by Alpha5-chain, Beta1-chain, and Gamma1-chain, is called "Laminin-511". Laminin-511 is recognized by alpha6beta1 integrin. Cell adhision, extension and proliferation is caused by many cell signalings through intergrin.

 

Laminin-511 E8 is the fragment of Laminin-511 obtained by enzyme digestion. Although this is fragment, it has binding ability to alpha6beta1 integrin. According to Miyazaki's report ( Nat. Commun. 3, 1236 (2012)), Laminin-511 E8 could be used for cell culture of ES cells and iPS cells.

 

Fig.1. Laminin 511-E8의 모식도 

 

To date, mouse feeder cells or matrigel have been used for the culture of iPS cells and ES cells. However, these cell culture substrates have a risk to bring xeno-components in the culture condition so this was said one of the problems for future medical and clinical application.

 
iMartix can be used for not only stem cells, but various cells, like epidermal cells, vascular endothelial cells. 

 

Basic Usage

1. Add 350uL of sterile tissue culture grade water to make 500ug/ml solution.

2. Dilute the solution with PBS (-). Coat dishes with 0.5ug/cm².

* The optimum coating concentration depends on cell lines from 0.1 to 1.5ug/cm².

** For example, when you use 6-well plate (9.6 cm²/well), add 10ul iMatrix-511 (500ug/ml) and 1.99ml PBS(-) for 1 well (2.4ug/ml, 2ml/well).

3. Incubate for 3 hours at Room temperature or over night at 4C.

4. Remove excess fluid from the coated surface.

5. Immediately plate the cells at desired density.

------------------------------------------------------------------------------------------------

*****When culturing ES/iPS cell, Feeder Free and Single cell subculture can be done using this product. *****

 

Example of the use of Laminin 511-E8

Laminin 511-E8 is not only the substrate of cell culture for epidermal cells and vascular endothelial cells, but also for iPS and ES cells.  Preparing for cell culture using Laminin 511-E8 is very simple.  Dilute it with PBS, and coat the petri dish or microscope slide by simple pouring it in.  Remove any excess solution and add medium.  Figure 2 shows the cultured epidermal cells and vascular endothelial cell using Laminin 511-E8.  Figure 2 (a) shows the cultured epidermal cell for both uncoated and coated with Laminin 511-E8.  It can be observed that the one coated with Laminin 511-E8 has far greater amount of cell adhesion than the uncoated one.  Figure 2 (b) shows the cultured vascular endothelial cell for both uncoated and coated with Laminin 511-E8.  The uncoated one show adhesion but most of the cells are still circular.  Meanwhile, the one coated with Laminin 511-E8 shows adhesion and the cells are not circular showing that the cells are spreading.  Thus by using Laminin 511-E8, it is possible to promote adhesion and spreading for various types of cells.

Fig.2. Human Epidemal Cells and Vascular Endothelical Cells cultured on iMartix-511. More efficient adhesion and extension can be seen when cultured on iMatrix-511.

 

It is known that handling hESC and hiPSC can be very difficult.  For example, when growing these types of cells, it must be divided into appropriate colony sizes.  If the cells completely dissociates, it will lead to the cell’s death.  on the other hand if the size of the colonies is not right the cells may differentiate.  Furthermore, standard culture substrate such as, feeder cells or Matrigel, are derived from mouse, which increases the risk caused by heterologous components, one of the main issues for using stem cells for medical treatments.  In light of these difficulties, findings from the collaborated research of Osaka University & Kyoto University that Laminin 511-E8 can also be used for iPS and ES cell culture, provided progress in resolving these issues, as using Laminin 511-E8 has been shown that many cells survive and continue to grow even if it get disassociated.  Also, because it is a recombinant protein the risk caused due to heterologous components are greatly decreased.   

 

Reference Literature 

1. Miner, JH and Yurchenco, PD Laminin functions in tissue morphogenesis. Annu. Rev. Cell Dev. Biol. 20, 255-284 (2004). 

2. Ido H, Harada K, Futaki S, Hayashi Y, Nishiuchi R, Natsuka Y, Li S, Wada Y, Combs AC, Ervasti JM and Sekiguchi K. Molecular dissection of the alpha-dystroglycan- and integrin-binding sites within the globular domain of human laminin-10. J. Biol. Chem. 279, 10946-10954 (2004). 

3. Ido H, Nakamura A, Kobayashi R, Ito S, Li S, Futaki S and Sekiguchi K. The requirement of the glutamic acid residue at the third position from the carboxyl termini of the laminin gamma chains in integrin binding by laminins. J. Biol. Chem. 282, 11144-11154 (2007). 

4. Taniguchi Y, Ido H, Sanzen N, Hayashi M, Sato-Nishiuchi R, Futaki S and Sekiguchi K. The C-terminal region of laminin beta chains modulates the integrin binding affinities of laminins. J. Biol. Chem. 284, 7820-7831 (2009). 

5. Miyazaki T, Futaki S, Suemori H, Taniguchi Y, Yamada M, Kawasaki M, Hayashi M, Kumagai H, Nakatsuji N, Sekiguchi K and Kawase E. Laminin E8 fragments support efficient adhesion and expansion of dissociated human pluripotent stem cells. Nat. Commun. 3, 1236 (2012). 

6. Nakagawa M, Taniguchi Y, Senda S, Takizawa N, Ichisaka T, Asano K, Morizane A, Doi D,  Takahashi J, Nishizawa M, Yoshida Y, Toyoda T, Osafune K, Sekiguchi K, Yamanaka S.  A novel efficient feeder-free culture system for the derivation of human induced pluripotent stem cells. SCIENTIFIC REPORTS | 4 : 3594 | DOI: 10.1038/srep03594 2014 Jan. 8

 

FOR RESEARCH USE onLY, NOT FOR USE IN DIAGNOSTIC PROCEDURES.

 

코아사이언스 coresciences nippi funakoshi 후나코시 한국 공식 대리점 distributor

iMatrix511_NIP_140708_CoreSciences.pdf

0.35MB

iMatrix511_NIP_140708_CoreSciences.pdf

0.35MB

iMatrix511_NIP_140708_CoreSciences.pdf

0.35MB