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干細(xì)胞組織消化手冊(cè)

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Worthington組織消化指南

引言

組織消化酶主要用于組織培養(yǎng)和細(xì)胞生物學(xué)研究的原代細(xì)胞的分離和收集。盡管這些消化酶已經(jīng)普遍應(yīng)用多年,但是對(duì)其組織消化和細(xì)胞收集的作用機(jī)制卻沒(méi)有全面的理解。  由此導(dǎo)致的結(jié)果便是選擇一種消化方法大多是隨機(jī)選擇或基于過(guò)去的經(jīng)驗(yàn),而不是在*理解消化原理的基礎(chǔ)上選擇的消化方案,以及如何根據(jù)具體實(shí)驗(yàn)?zāi)康男拚桨敢垣@得更好的實(shí)驗(yàn)結(jié)果。

細(xì)胞分離程序的目的是zui大程度的獲得功能活性的單個(gè)細(xì)胞,而影響結(jié)果的原因有很多,包括(不限于)以下:

1、組織類(lèi)型

2、物種來(lái)源

3、動(dòng)物年齡

4、遺傳改造情況(如knockouts

5、所用的消化培養(yǎng)液

6、所用的酶

7、消化酶中的雜質(zhì)

8、所用消化酶的工作濃度

9、溫度

10、孵育時(shí)間

通常前四個(gè)原因沒(méi)辦法選擇,而剩下的原因一般由經(jīng)驗(yàn)決定以獲取zui適實(shí)驗(yàn)結(jié)果。

通過(guò)查閱文獻(xiàn)來(lái)選擇*組織消化酶和zui適消化條件的研究人員通常被互相矛盾的實(shí)驗(yàn)數(shù)據(jù)搞得一頭霧水。這些巨大差異來(lái)源于組織胞外基質(zhì)的復(fù)雜而動(dòng)態(tài)的特性,以及過(guò)去常用的粗制、未提純的組織分離酶。另外,組織胞外基質(zhì)由各種蛋白、糖蛋白、脂類(lèi)和糖脂組成,這些成分的組成比例在不同物種、不同組織、甚至不同發(fā)育(年齡)之間存在明顯的差異,這些因素都極大的影響了消化方案的制定。目前通常使用的粗制酶,如鏈霉蛋白酶、NF1:250、以及含有幾種其他不同濃度蛋白酶的膠原酶、以及各種多糖酶、核酶和脂酶。

本指南總結(jié)了目前常用組織消化酶的zui適消化和細(xì)胞收集知識(shí)和方法,描述了標(biāo)準(zhǔn)的實(shí)驗(yàn)步驟, 便于制定一個(gè)*的細(xì)胞分離方案,同時(shí)列舉了一些相關(guān)的參考文獻(xiàn)。

下表列出多種組織的適宜消化酶(含參考文獻(xiàn),以組織類(lèi)型和物種分類(lèi),詳見(jiàn)鏈接)

脂肪組織

腎上腺組織

骨組織

腦組織

軟骨組織

結(jié)腸組織

內(nèi)皮組織

上皮組織

心臟

淋巴結(jié)

乳腺

其它組織

其他組織

肌組織

神經(jīng)組織

胰腺

腮腺

垂體

前列腺

生殖

動(dòng)物鱗片

皮膚

脾臟

干細(xì)胞

胸腺

甲狀腺/甲狀旁腺

扁桃體

腫瘤

 

 

 

注:使用Worthington公司消化酶的文獻(xiàn)不限于上表鏈接中所列,一般而言,Worthington公司的組織消化酶可用于大多組織類(lèi)型的消化。

 

干細(xì)胞組織消化手冊(cè)

 

物種詳細(xì)來(lái)源細(xì)胞名稱(chēng)消化用酶培養(yǎng)基參考文獻(xiàn)
鳥(niǎo)Adipose derived stem cellsCollagenase Type 1: 0.1%DMEMTandon, N., Goh, B., Marsano, A., Chao, P., Montouri-Sorrentino, C., Gimble, J. and Vunjak-Novakovic, G.: Alignment and Elongation of Human Adipose-Derived Stem Cells in Response to Direct-Current Electrical Stimulation., Conf Proc IEEE Eng Med Biol Soc 2009, 6517-21, 2009
Amniotic membrane MSCTrypsin: 0.25%LG-DMEMPark, S., Seo, M., Kim, H. and Kang, K.: Isolation and Characterization of Canine Amniotic Membrane-Derived Multipotent Stem Cells., PLoS ONE 7, e44693, 2012
Collagenase Type 1: 0.2%
犬, 20-25 公斤Adipose stem cellsCollagenase: see referenceMedia-199Fischer, L., McIlhenny, S., Tulenko, T., Golesorkhi, N., Zhang, P., Larson, R., Lombardi, J., Shapiro, I. and DiMuzio, P.: Endothelial Differentiation of Adipose-Derived Stem Cells: Effects of Endothelial Cell Growth Supplement and Shear Force., J Surg Res 152, 157, 2009
Adipose derived stem cellsCollagenase Type 1: 0.1%PBSVidal, M., Robinson, S., Lopez, M., Paulsen, D., Borkhsenious, O., Johnson, J., Moore, R. and Gimble, J.: Comparison of Chondrogenic Potential in Equine Mesenchymal Stromal Cells Derived from Adipose Tissue and Bone Marrow., Vet Surg 37, 713, 2008
馬, 1-5 歲Adipose derived stem cellsCollagenase Type 1: 0.1%PBSVidal, M., Kilroy, G., Lopez, M., Johnson, J., Moore, R. and Gimble, J.: Characterization of Equine Adipose Tissue-Derived Stromal Cells: Adipogenic and Osteogenic Capacity and Comparison with Bone Marrow-Derived Mesenchymal Stromal Cells., Vet Surg 36, 613, 2007
人類(lèi)人類(lèi)Mesenchymal Stromal CellsCollagenase Type 4: 0.2%PBSChoudhery,M, Badowski, M, Muise, A and Harris, D: Effect of Mild Heat Stress on the Proliferative and Differentiative Ability of Human Mesenchymal Stromal Cells., Cytotherapy17, 359-68, 2015
人類(lèi)人類(lèi)Adipose derived stem cellsCollagenase Type 2: 0.1%HBSSSatish, L., Krill-Burger, J., Gallo, P., Etages, S., Liu, F., Philips, B., Ravuri, S., Marra, K., LaFramboise, W., Kathju, S. and Rubin, J.: Expression Analysis of Human Adipose-Derived Stem Cells During In Vitro Differentiation to an Adipocyte Lineage., BMC Med Genomics 8, 41, 2015
人類(lèi)人類(lèi)Dental mesenchymal stemCollagenase Type 1: 0.3%DMEMWu, Z., Wang, J., Dong, R., Wang, L., Fan, Z., Liu, D. and Wang, S.: Depletion of MEIS2 Inhibits Osteogenic Differentiation Potential of Human Dental Stem Cells., Int J Clin Exp Med8, 7220-30, 2015
Neutral Protease: 0.4%
人類(lèi)人類(lèi)Decidua Mesenchymal StemTrypsin: 0.25%PBSKusuma, G., Menicanin, D., Gronthos, S., Manuelpillai, U., Abumaree, M., Pertile, M., Brennecke, S. and Kalionis, B.: Ectopic Bone Formation by Mesenchymal Stem Cells Derived from Human Term Placenta and the Decidua., PLoS ONE 10, e0141246, 2015
Collagenase Type 1: 1.0%
Deoxyribonuclease I: 0.005%
人類(lèi)人類(lèi), 成年人Adipose derived mesenchymal stemCollagenase Type 2: 0.1%see referenceAl-Saqi, S., Saliem, M., Asikainen, S., Quezada, H., Ekblad, A., Hovatta, O., Le Blanc, K., Jonasson, A. and Gotherstrom, C.: Defined Serum-Free Media for In Vitro Expansion of Adipose-Derived Mesenchymal Stem Cells., Cytotherapy 16, 915, 2014
人類(lèi)人類(lèi), 成年人Adipose derived stem cellsCollagenase Type 1: 0.15%DMEMKoellensperger, E., Bollinger, N., Dexheimer, V., Gramley, F., Germann, G. and Leimer, U.: Choosing the Right Type of Serum for Different Applications of Human Adipose Tissue-Derived Stem Cells: Influence on Proliferation and Differentiation Abilities., Cytotherapy 16, 789, 2014
人類(lèi)人類(lèi)Umbilical mesenchymal stemCollagenase Type 2: 0.1%DMEMHang, H., Yu, Y., Wu, N., Huang, Q., Xia, Q. and Bian, J.: Induction of Highly Functional Hepatocytes from Human Umbilical Cord Mesenchymal Stem Cells by HNF4偽 Transduction.,PLoS ONE 9, e104133, 2014
Trypsin: 0.25%
人類(lèi)人類(lèi)Cardiac stemCollagenase Type 2: 0.25%Joklik modified Eagle抯Avolio, E., Gianfranceschi, G., Cesselli, D., Caragnano, A., Athanasakis, E., Katare, R., Meloni, M., Palma, A., Barchiesi, A., Vascotto, C., Toffoletto, B., Mazzega, E., Finato, N., Aresu, G. and Livi, U.: Ex vivo Molecular Rejuvenation Improves the Therapeutic Activity of Senescent Human Cardiac Stem Cells in a Mouse Model of Myocardial Infarction., Stem Cells 32, 2373, 2014
人類(lèi)人類(lèi)Adipose stromal stemCollagenase animal free: 200 u/mlDMEM/Hams F-12Carvalho, P., Gimble, J., Dias, I., Gomes, M. and Reis, R.: Xenofree Enzymatic Products for the Isolation of Human Adipose-Derived Stromal/Stem Cells., Tiss Eng 19, 473-8, 2013
人類(lèi)人類(lèi), 成年人Epidermal stem cellsNeutral Protease: 2 u/mlDMEMNagel, S., Rohr, F., Weber, C., Kier, J., Siemers, F., Kruse, C., Danner, S., Brandenburger, M. and Matthiessen, A.: Multipotent Nestin-Positive Stem Cells Reside in the Stroma of Human Eccrine and Apocrine Sweat Glands and can be Propagated Robustly In Vitro., PLoS ONE 8, e78365, 2013
Trypsin: 2.5%
人類(lèi)人類(lèi)Dental pulp stemCollagenase Type 1: 0.3%DMEM/F12Bonnamain, V., Thinard, R., Sergent-Tanguy, S., Huet, P., Bienvenu, G., Naveilhan, P., Farges, J. and Alliot-Licht, B.: Human Dental Pulp Stem Cells Cultured in Serum-Free Supplemented Medium., Front Physiol 4, 357, 2013
Neutral Protease: 0.4%
人類(lèi)人類(lèi)Umbilical Cord Mesenchymal StemCollagenase: 0.1-0.2%DMEMHan, Y., Tao, R., Sun, T., Chai, J., Xu, G. and Liu, J.: Optimization of Human Umbilical Cord Mesenchymal Stem Cell Isolation and Culture Methods., Cytotechnology 65, 819, 2013
Trypsin: 0.1%
人類(lèi)人類(lèi)Dental pulp derived stem cellCollagenase Type 1: 0.3%DMEMSakai, K., Yamamoto, A., Matsubara, K., Nakamura, S., Naruse, M., Yamagata, M., Sakamoto, K., Tauchi, R., Wakao, N., Imagama, S., Hibi, H., Kadomatsu, K., Ishiguro, N. and Ueda, M: Human Dental Pulp-Derived Stem Cells Promote Locomotor Recovery After Complete Transection of the Rat Spinal Cord by Multiple Neuro-Regenerative Mechanisms., J Clin Invest 122, 80, 2012
Neutral Protease: 0.4%
人類(lèi)人類(lèi)Adipose-derived stem cellsCollagenase Type 1: 0.1%PBSCervelli, V., Scioli, M., Gentile, P., Doldo, E., Bonanno, E., Spagnoli, L. and Orlandi, A.:Plaet-Rich Plasma Greatly Potentiates Insulin-Induced Adipogenic Differentiation of Human Adipose-Derived Stem Cells Through a Serine/Threonine Kinase Akt-dependent Mechanism and Promotes Clinical Fat Graft Maintenance., Stem Cells Transl Med 1, 206-20, 2012
人類(lèi)人類(lèi)Mesenchymal stemCollagenase Type 1: 0.4%DMEM/F12Shalini, V., Pratheep, S., Muhammad, A., Sharmili, V., Elizabeth, G. and Rajesh, R.: Generation and Characterisation of Human Mesenchymal Stem Cells Derived From Umbilical Cord and Placenta, Regenerative Research 1, 48, 2012
Deoxyribonuclease I: 0.01%
人類(lèi)人類(lèi)Wharton jelly mesenchymal stemCollagenase Type 2: 0.1%DMEMSalehinejad, P., Alitheen, N., Ali, A., Omar, A., Mohit, M., Janzamin, E., Samani, F., Torshizi, Z. and Nematollahi-Mahani, S.: Comparison of Different Methods for the Isolation of Mesenchymal Stem Cells from Human Umbilical Cord Wharton's Jelly., In Vitro Cell Dev Biol Anim 48, 75, 2012
Trypsin: 0.25%
人類(lèi)人類(lèi)Placental Mesenchymal StemCollagenase Type 2: 0.4%DMEMVellasamy, S., Sandrasaigaran, P., Vidyadaran, S., George, E. and Ramasamy, R.:Isolation and Characterisation of Mesenchymal Stem cells Derived from Human Placenta Tissue., World J Stem Cells 4, 53-61, 2012
Deoxyribonuclease I: 0.01%
人類(lèi)人類(lèi), 男性 40-60 yearsAdipose derived stem cellsCollagenase: 0.25%PBSBlasi, A., Martino, C., Balducci, L., Saldarelli, M., Soleti, A., Navone, S., Canzi, L., Cristini, S., Invernici, G., Parati, E. and Alessandri, G.: Dermal Fibroblasts Display Similar Phenotypic and Differentiation Capacity to Fat-Derived Mesenchymal Stem Cells, but Differ in Anti-Inflammatory and Angiogenic Potential, Vasc Cell 3, 5, 2011
Deoxyribonuclease I: 0.002%
人類(lèi)人類(lèi)Umbilical cord stromal stemCollagenase Type 4: 0.08%DMEMFarias, V., Linares-Fernandez, J., Penalver, J., Paya Colmenero, J., Ferron, G., Duran, E., Fernandez, R., Olivares, E., O'Valle, F., Puertas, A., Oliver, F. and Ruiz de Almodovar, J.:Human Umbilical Cord Stromal Stem Cell Express CD10 and Exert Contractile Properties.,Placenta 32, 86, 2011
Neutral Protease: 0.138%
Hyaluronidase: 0.02%
人類(lèi)人類(lèi)Vascular wall-resident multipotent stem cellsCollagenase Type 2: 0.2%see referenceKlein, D., Weisshardt, P., Kleff, V., Jastrow, H., Jakob, H., and Ergun, S.: Vascular Wall-Resident CD44+ Multipotent Stem Cells Vive Rise to Pericytes and Smooth Muscle Cells and Contribute to New Vessel Maturation., PLoS ONE 6, e20540, 2011
Elastase: 5 u/ml
人類(lèi)人類(lèi)Adipose derived stemCollagenase Type 1: 0.1%DMEM/Ham's F-12Yu, G. Floyd, ZE., Wu, X., Halvorsen, Y. and Gimble, J.: Isolation of Human Adipose-Derived Stem Cells from Lipoaspirates., Methods Mol Biol 702, 17-27, 2011
人類(lèi)人類(lèi)Glioma stem cellsPDS kit: per instructionsneurobasal mediumHjelmeland, A., Wu, Q., Wickman, S., Eyler, C., Heddleston, J., Shi, Q., Lathia, J., Macswords, J., Lee, J., McLendon, R. and Rich, J.: Targeting A20 Decreases Glioma Stem Cell Survival and Tumor Growth., PLoS Biol 8, e1000319, 2010
人類(lèi)人類(lèi)Dental pulp and apical papilla stem cellsCollagenase Type 1: 0.3%MEMHuang, G., Yamaza, T., Shea, L., Djouad, F., Kuhn, N., Tuan, R. and Shi, S.:Stem/Progenitor Cell-Mediated De Novo Regeneration of Dental Pulp with Newly Deposited Continuous Layer of Dentin in an In Vivo Model., Tissue Eng Part A 16, 605, 2010
Neutral Protease: 0.4%
人類(lèi)人類(lèi)Corneal stromal stemNeutral Protease: 1.2 u/mlDMEMDu, Y., Roh, D., Funderburgh, M., Mann, M., Marra, K., Rubin, J., Li, X. and Funderburgh, J.: Adipose-Derived Stem Cells Differentiate to Keratocytes In Vitro., Mol Vis 16, 2680, 2010
Collagenase: 0.1%
人類(lèi)人類(lèi)Adipose tissue-derived stemCollagenase Type 2: 1.0%DMEM/F12Tan, H., DeFail, A., Rubin, J., Chu, C. and Marra, K.: Novel Multiarm PEG-Based Hydrogels for Tissue Engineering., J Biomed Mater Res 92, 979, 2010
人類(lèi)人類(lèi)Umbilical cord mesenchymalHyaluronidase: 0.05%DMEMZeddou, M., Briquet, A., Relic, B., Josse, C., Malaise, M., Gothot, A., Lechanteur, C. and Beguin, Y.: The Umbilical Cord Matrix is a Better Source of Mesenchymal Stem Cells (MSC) Than the Umbilical Cord Blood., Cell Biol Int 34, 693-701, 2010
Collagenase: 0.08%
人類(lèi)人類(lèi)Adipose stem cellsCollagenase Type 1: 0.1%DMEMHareendran, S., Sathishkumar, S., Abbas, S., Mackay, A. and Rajan, P.: A Novel Composition for the Culture of Human Adipose Stem Cells which Includes Complement C3.,Cytotechnology 62, 389-402, 2010
人類(lèi)人類(lèi), 40-65 yearAdult human adipose stem cellsCollagenase Type 2: 0.075%DMEMSun, N., Panetta, N., Gupta, D., Wilson, K., Lee, A., Jia, F., Hu, S., Cherry, A., Robbins, R., Longaker, M. and Wu, J.: Feeder-Free Derivation of Induced Pluripotent Stem Cells from Adult Human Adipose Stem Cells., Proc Natl Acad Sci U S A 106, 15720, 2009
人類(lèi)人類(lèi)Filum terminale neural progenitorTrypsin: see referenceDMEM/F-12Varghese, M., Olstorn, H., Berg-Johnsen, J., Moe, M., Murrell, W. and Langmoen, I.:Isolation of Human Multipotent Neural Progenitors from Adult Filum Terminale., Stem Cells Dev 18, 603, 2009
人類(lèi)人類(lèi)skin and keloid progenitor cellsCollagenase Type 1: 0.4%PBSZhang, Q., Yamaza, T., Kelly, AP, Shi, S., Wang, S., Brown, J., Wang, L., French, S., Shi, S. and Le, A.: Tumor-like Stem Cells Derived from Human Keloid are Governed by the Inflammatory Niche Driven by IL-17/IL-6 Axis., PLoS ONE 4, e7798, 2009
Neutral Protease: 0.3%
人類(lèi)人類(lèi), 男性Spermatogonial stem cellsCollagenase: 1%DMEMKossack, N., Meneses, J., Shefi, S., Nguyen, H., Chavez, S., Nicholas, C., Gromoll, J., Turek, P. and Reijo-Pera, R.: Isolation and Characterization of Pluripotent Human Spermatogonial Stem Cell-Derived Cells., Stem Cells , , 2008
Deoxyribonuclease I: 0.22%
Trypsin: 0.4%
人類(lèi)人類(lèi)Muscle derived multiprogenitor cellsCollagenase Type 2: 0.05%DMEMNesti, L., Jackson, W., Shanti, R., Koehler, S., Aragon, A., Bailey, J., Sracic, M., Freedman, B., Giuliani, J. and Tuan, R.: Differentiation Potential of Multipotent Progenitor Cells Derived from War-Traumatized Muscle Tissue., J Bone Joint Surg Am 90, 2390, 2008
人類(lèi)人類(lèi)Adipose derived stem cellsCollagenase Type 1: 0.075%DMEMJeong, J.: Adipose Stem Cells as a Clinically Available and Effective Source of Adult Stem Cell Therapy, Int J Stem Cells 1, 43, 2008
人類(lèi)人類(lèi)Hepatic stem cells and heptoblastsCollagenase Type 4: 0.014-0.06%variousWauthier, E., Schmelzer, E., Turner, W., Zhang, L., LeCluyse, E., Ruiz, J., Turner, R., Furth, M., Kubota, H., Lozoya, O., Barbier, C., McClelland, R., Yao, H., Moss, N., Bruce, A., Ludlow, J. and Reid, L.: Hepatic Stem Cells and Hepatoblasts: Identification, Isolation, and Ex Vivo Maintenance., Methods Cell Biol 86, 137, 2008
人類(lèi)人類(lèi)Umbilical mesenchymal stemCollagenase: 0.1%DMEMSecco, M., Zucconi, E., Vieira, N., Fogaca, L.,Cerqueira, A., Carvalho, M., Jazedje, T., Okamoto, O., Muotri, A. and Zatz, M.: Multipotent Stem Cells from Umbilical Cord: Cord is Richer Than Blood!, Stem Cells 26, 146, 2008
人類(lèi)人類(lèi)Stem CellsCollagenase: 280 u/mlD-PBSPilgaard, L., Lund, P., Rasmussen, J., Fink, T. and Zachar, V.: Comparative Analysis of Highly Defined Proteases for the Isolation of Adipose Tissue-Derived Stem Cells., Regen Med3, 705-15, 2008
人類(lèi)人類(lèi)Bone marrow derived MSCTrypsin: 0.05%DMEMWelter, J., Solchaga, L.and Penick, K.: Simplification of Aggregate Culture of Human Mesenchymal Stem Cells as a Chondrogenic Screening Assay, Biotechniques 42, 732, 2007
Papain: 0.0025%
人類(lèi)人類(lèi), 8-12 yearTendon stem/progenitorCollagenase Type 1: 0.3%DMEMBi Yanming, Ehirchiou Driss, Kilts Tina M, Inkson Colette A, Embree Mildred C, Sonoyama Wataru, Li Li, Leet Arabella I, Seo Byoung-Moo, Zhang Li, Shi Songtao, Young Marian F:Identification of tendon stem/progenitor cells and the role of the extracellular matrix in their niche, Nat Med 13, 1219-27, 2007
Neutral Protease: 0.4%
人類(lèi)人類(lèi), 男性Adipose derived adult stem cellsCollagenase Type 1: 0.1%DMEMLei, L., Liao, W., Sheng, P., Fu, M., He, A. and Huang, G.: Biological Character of Human Adipose-Derived Adult Stem Cells and Influence of Donor Age on Cell Replication in Culture.,Sci China C Life Sci 50, 320, 2007
人類(lèi)人類(lèi)Pancreatic cancer stem cellsCollagenase Type 4: 200 u/mlmedium 199Li, C., Heidt, D., Dalerba, P., Burant, C., Zhang, L., Adsay, V., Wicha, M., Clarke, M. and Simeone, D.: Identification of Pancreatic Cancer Stem Cells., Cancer Res 67, 1030, 2007
人類(lèi)人類(lèi), 成年人Adipose-derived adult stemCollagenase Type 1: 0.1%DMEM/F-12 Ham'sMitchell James B, McIntosh Kevin, Zvonic Sanjin, Garrett Sara, Floyd Z Elizabeth, Kloster Amy, Di Halvorsen Yuan, Storms Robert W, Goh Brian, Kilroy Gail, Wu Xiying, Gimble Jeffrey M: Immunophenotype of human adipose-derived cells: temporal changes in stromal-associated and stem cell-associated markers, Stem Cells 24, 376-85, 2006
人類(lèi)人類(lèi)Mesenchymal stemCollagenase Type 1: 0.075%DMEMKern Susanne, Eichler Hermann, Stoeve Johannes, Kluter Harald, Bieback Karen:Comparative analysis of mesenchymal stem cells from bone marrow, umbilical cord blood, or adipose tissue, Stem Cells 24, 1294-301, 2006
人類(lèi)人類(lèi)Placental mesenchymal stemCollagenase Type 2: 270 u/mlMEMPortmann-Lanz CBettina, Schoeberlein Andreina, Huber Alexander, Sager Ruth, 男性k Antoine, Holzgreve Wolfgang, Surbek DanielV: Placental mesenchymal stem cells as potential autologous graft for pre- and perinatal neuroregeneration, Am J Obstet Gynecol194, 664-73, 2006
Neutral Protease: 2.4 u/ml
人類(lèi)人類(lèi)Stromal stem cellsCollagenase Type 1: 0.2%HBSSBoquest Andrew C, Shahdadfar Aboulghassem, Brinchmann Jan E, Collas Philippe:Isolation of stromal stem cells from human adipose tissue, Methods Mol Biol 325, 35-46, 2006
人類(lèi)人類(lèi), 女性Adipose derived adult stem cellsCollagenase Type 1: 0.1%DMEM/F-12Guilak, F., Lott, K., Awad, H., Cao, Q., Hicok, K., Fermor, B. and Gimble, J.: Clonal Analysis of the Differentiation Potential of Human Adipose-Derived Adult Stem Cells., J Cell Physiol206, 229, 2006
人類(lèi)人類(lèi)Adult stem cellsCollagenase Type 1: 0.1%PBSDevireddy, R., Thirumala, S. and Gimble, J.: Cellular Response of Adipose Derived Passage-4 Adult Stem Cells to Freezing Stress., J Biomech Eng 127, 1081, 2005
人類(lèi)人類(lèi), 女性Adipose derived adult stem cellsCollagenase Type 1: 0.1%DMEM-Ham's F-12Aust, L., Devlin, B., Foster, S., Halvorsen, Y., Hicok, K., du Laney, T., Sen, A., Willingmyre, G. and Gimble, J.: Yield of Human Adipose-Derived Adult Stem Cells from Liposuction Aspirates., Cytometry 6, 7-14, 2004
人類(lèi)人類(lèi), 成年人, 男性Human epidermal keratinocyte stem cellsNeutral Protease: 0.5%DMEMPapini S, Cecchetti D, Campani D, Fitzgerald W, Grivel J, Chen S, Margolis L, Revolla R.: Isolation and Clonal Analysis of Human Epidermal Keratinocyte Stem Cells in Long-term Culture, Stem Cells 21, 481, 2003
人類(lèi)人類(lèi)Umbilical vein mesenchymal stem cellsCollagenase: 1%PBSCovas DT, Siufi JL C, Silva AR L, Orellana MD: Isolation and culture of umbilical vein mesenchymal stem cells, Braz J Med Biol Res 36, 1179-83, 2003
人類(lèi)人類(lèi), 7-8 歲Stem cells Human exfoliated deciduous teethCollagenase Type 1: 0.3%see referenceMiura, M., Gronthos, S, Zhao, M, Lu, B, Fisher, L, Robey, P, and Shi, S.: SHED: Stem Cells from Human Exfoliated Deciduous Teeth, Proc Natl Acad Sci U S A 100, 5807, 2003
Neutral Protease: 0.4%
人類(lèi)人類(lèi), 胎兒Epithelial progenitorCollagenase: 0.03%DMEMMalhi, H., Irani, A., Gagandeep, S. and Gupta, S.: Isolation of Human Progenitor Liver Epithelial Cells with Extensive Replication Capacity and Differentiation into Mature Hepatocytes., J Cell Sci 115, 2679, 2002
人類(lèi)人類(lèi), 成年人Human skin mast cellsCollagenase Type 2: 0.15%HBSSKambe, N, Kambe, M, Kochan, J, and Schwartz ,L.: Human Skin-derived Mast Cells Can Proliferate While Retaining Their Characteristic Functional and Protease Phenotypes, Blood 97, 2045, 2001
Hyaluronidase: 0.07%
Deoxyribonuclease I: 0.03%
人類(lèi)人類(lèi)Central nervous system stemCollagenase: 0.1%HBSSUchida, N., Buck, D., He, D., Reitsma, M., Masek, M., Phan, T., Tsukamoto, A., Gage, F. and Weissman, I.: Direct Isolation of Human Central Nervous System Stem Cells., Proc Natl Acad Sci U S A 97, 14720, 2000
Hyaluronidase: 0.1%
人類(lèi)人類(lèi)Embryonic stemNeutral Protease: 1%DMEMThomson JA, Itskovitz-Eldor J, Shapiro SS, Waknitz MA, Swiergiel JJ, Marshall VS, Jones JM: Embryonic stem cell lines derived from human blastocysts, Science 282, 1145-7, 1998
Collagenase Type 4: 0.1%
人類(lèi)人類(lèi)Muscle-derived stem cellsTrypsin: 0.25%DMEM/F12Alessandri Giulio, Pagano Stefano, Bez Alessandra, Benetti Anna, Pozzi Stefano, Iannolo Gioacchin, Baronio Manuela, Invernici Gloria, Caruso Arnaldo, Muneretto Claudio, Bisleri Gianluigi, Parati Eugenio: Isolation and culture of human muscle-derived stem cells able to differentiate into myogenic and neurogenic cell lineages, Lancet 364, 1872-83,
Embryonic stemCollagenase Type 4: 0.08%DMEMChen, S., Revolla, R., Papini, S., Michelini, M., Fitzgerald, W., Zimmerberg, J.,and Margolis, L.: Multilineage Differentiation of Rhesus Monkey Embryonic Stem cells in Three-dimensional Culture Systems, Stem Cells 21(3), 281, 2003
Baboons, 1歲, 7歲, 14 歲Primate spermatogonialCollagenase Type 2: 0.1%DMEMNagano, M., McCarrey, J., and Brinster, R.: Primate Spermatogonial Stem Cells Colonize Mouse Testes, Biol Reprod 64, 1409, 2001
Trypsin: 0.05%
Deoxyribonuclease I: 0.1%
小鼠小鼠, 3-4 周Cardiac progenitorCollagenase Type 1: 0.1%DMEMWang, H., Chen, H., Feng, B., Wang, X., He, X., Hu, R., Yin, M., Wang, W., Fu, W. and Xu, Z.: Isolation and Characterization of a Sca-1+/CD31- Progenitor Cell Lineage Derived From Mouse Heart Tissue., BMC Biotechnol 14, 75, 2014
小鼠小鼠, P4Enteric neural crest progenitorsNeutral Protease: 0.5%DMEM/F12Hotta, R., Stamp, L., Foong, J., McConnell, S., Bergner, A., Anderson, R., Enomoto, H., Newgreen, D., Obermayr, F., Furness, J. and Young, H.: Transplanted Progenitors Generate Functional Enteric Neurons in the Postnatal Colon., J Clin Invest 123, 1182, 2013
Collagenase animal free: 0.05%
小鼠小鼠Epithelial StemTrypsin: 0.25%HBSSJensen, U., Ghazizadeh, S. and Owens, D.: Isolation and Characterization of Cutaneous Epithelial Stem Cells., Methods Mol Biol 989, 61-9, 2013
Collagenase Type 1: 0.4%
小鼠小鼠, 10 周Synovial mesenchymalCollagenase: 0.1%DMEMFutami, I., Ishijima, M., Kaneko, H., Tsuji, K., Ichikawa-Tomikawa, N., Sadatsuki, R., Muneta, T., Arikawa-Hirasawa, E., Sekiya, I. and Kaneko, K.: Isolation and Characterization of Multipotential Mesenchymal Cells from the Mouse Synovium., PLoS ONE7, e45517, 2012
Deoxyribonuclease I: 0.005%
小鼠小鼠, 6-8 周Neural stem cellsPapain: 0.01%DMEM/F12Bracko, O., Singer, T., Aigner, S., Knobloch, M., Winner, B., Ray, J., Clemenson, G., Suh, H., Couillard-Despres, S., Aigner, L., Gage, F. and Jessberger, S.: Gene Expression Profiling of Neural Stem Cells and their Neuronal Progeny Reveals IGF2 as a Regulator of Adult Hippocampal Neurogenesis., J Neurosci 32, 3376-87, 2012
Neutral Protease: 0.1%
Deoxyribonuclease I: 0.01%
小鼠小鼠, 8-10 周Adipose derived stemCollagenase Type 1: 0.025%HBSSSugii, S., Kida, Y., Berggren, W. and Evans, R.: Feeder-Dependent and Feeder-Independent iPS Cell Derivation from Human and Mouse Adipose Stem Cells., Nat Protoc 6, 346, 2011
小鼠小鼠Spleen, bone marrow endothelialCollagenase Type 4: 0.3-1.0%PBSShi, C., Jia, T., Mendez-Ferrer, S., Hohl, T., Serbina, N., Lipuma, L., Leiner, I., Li, M., Frenette, P. and Pamer, E.: Bone Marrow Mesenchymal Stem and Progenitor Cells Induce Monocyte Emigration in Response to Circulating Toll-Like Receptor Ligands., Immunity 34, 590, 2011
Deoxyribonuclease I: 20 u/ml
小鼠小鼠, 8-10 周Lung mesenchymal stemCollagenase Type 2: 0.2%HBSSChow, K., Jun, D., Helm, K., Wagner, D. and Majka, S.: Isolation & Characterization of Hoechst(low) CD45(negative) Mouse Lung Mesenchymal Stem Cells., J Vis Exp 56, e3159, 2011
小鼠小鼠, 新生的Dental Pulp Stem CellsCollagenase Type 4: 0.2%PBSJanebodin, K., Horst, O., Ieronimakis, N., Balasundaram, G., Reesukumal, K., Pratumvinit, B. and Reyes, M.: Isolation and Characterization of Neural Crest-Derived Stem Cells from Dental Pulp of Neonatal Mice., PLoS ONE 6, e27526, 2011
Neutral Protease: 1.2 u/ml
小鼠小鼠Stem and progenitorCollagenase Type 2: 0.2%HBSSHan, J., Koh, Y., Moon, H., Ryoo, H., Cho, C., Kim, I. and Koh, G.: Adipose Tissue is an Extramedullary Reservoir for Functional Hematopoietic Stem and Progenitor Cells., Blood 115, 957, 2010
小鼠小鼠, 6-8 周Bone marrow mesenchymal stemCollagenase Type 1: 0.25%RPMI 1640Xu, S., De Becker, A., Van Camp, B., Vanderkerken, K. and Van Riet, I.: An Improved Harvest and In Vitro Expansion Protocol for Murine Bone Marrow-Derived Mesenchymal Stem Cells., J Biomed Biotechnol 2010, 105940, 2010
小鼠小鼠, 成年人Adult neural stemTrypsin: 0.05%DMEMDeleyrolle, L. and Reynolds, B.: Isolation, Expansion, and Differentiation of Adult Mammalian Neural Stem and Progenitor Cells using the Neurosphere Assay., Methods Mol Biol 549, 91, 2009
小鼠小鼠, 6-8 周Prostate epithelial/stemCollagenase Type 2: 0.5%HBSSBurger, P., Gupta, R., Xiong, X., Ontiveros, C., Salm, S., Moscali, D. and Wilson, E.:High Aldehyde Dehydrogenase Activity: A Novel Functional Marker of Murine Prostate Stem/Progenitor Cells., Stem Cells 27, 2220-8, 2009
Trypsin: 0.05%
小鼠小鼠Adipose tissue-derived stemCollagenase Type 1: 0.075%Modified EaglesCho, K., Park, H., Park, H., Jung, J., Jeon, S., Kim, Y. and Roh, H.: IFATS Collection: Immunomodulatory Effects of Adipose Tissue-Derived Stem Cells in an Allergic Rhinitis Mouse Model., Stem Cells 27, 259-65, 2009
小鼠小鼠Germ cellsCollagenase Type 1: 100 u/mlHBSSBreault, D., Min, I., Carlone, D., Farilla, L., Ambruzs, D., Henderson, D., Algra, S., Montgomery, R., Wagers, A. and Hole, N.: Generation of mTert-GFP Mice as a Model to Identify and Study Tissue Progenitor Cells., Proc Natl Acad Sci U S A 105, 10420, 2008
小鼠小鼠Neural progenitor cellPapain: 10 ul/mlPBSHutton, S. and Pevny, L.: Isolation, Culture, and Differentiation of Progenitor Cells from the Central Nervous System., Cold Spring Harb. Protoc. 11, 5077, 2008
小鼠小鼠, 3-6 周Ear mesenchymal stemCollagenase Type 1: 0.2%DMEM/F12Staszkiewicz, J., Gimble, J., Manuel, J. and Gawronska-Kozak, B.: IFATS Collection: Stem Cell Antigen-1-Positive Ear Mesenchymal Stem Cells Display Enhanced Adipogenic Potential.,Stem Cells 26, 2666, 2008
小鼠小鼠, 胚胎HES-BC cellsTrypsin: 0.05%DMEMLu Shi-Jiang, Feng Qiang, Caballero Sergio, Chen Yu, Moore Malcolm A, Grant Maria B, Lanza Robert: Generation of functional hemangioblasts from human embryonic stem cells,Nat Methods 4, 501-9, 2007
小鼠小鼠, 6-8 周Tendon stem/progenitorCollagenase Type 1: 0.3%DMEMBi Yanming, Ehirchiou Driss, Kilts Tina M, Inkson Colette A, Embree Mildred C, Sonoyama Wataru, Li Li, Leet Arabella I, Seo Byoung-Moo, Zhang Li, Shi Songtao, Young Marian F:Identification of tendon stem/progenitor cells and the role of the extracellular matrix in their niche, Nat Med 13, 1219-27, 2007
Neutral Protease: 0.4%
小鼠小鼠Liver epithelial progenitor cellsCollagenase Type 4: 0.1%DMEMLi Wen-Lin, Su Juan, Yao Yu-Cheng, Tao Xin-Rong, Yan Yong-Bi, Yu Hong-Yu, Wang Xin-Min, Li Jian-Xiu, Yang Yong-Ji, Lau Joseph T Y, Hu Yi-Ping: Isolation and characterization of bipotent liver progenitor cells from adult mouse, Stem Cells 24, 322-32, 2006
Deoxyribonuclease I: 0.05%
小鼠小鼠, 6 周Adipose mesenchymal stemCollagenase: 0.2%PBSDi Rocco Giuliana, Iachininoto Maria Grazia, Tritarelli Alessandra, Straino Stefania, Zacheo Antonella, Germani Antonia, Crea Filippo, Capogrossi Maurizio C: Myogenic potential of adipose-tissue-derived cells, J Cell Sci 119, 2945-52, 2006
小鼠小鼠Neural stem cellsPapain: see referenceDMEM/F-12Meletis, K., Wirta, V., Hede, S., Nister, M., Lundeberg, J. abd Frisen, J.: p53 Suppresses the Self-Renewal of Adult Neural Stem Cells., Development 133, 363, 2006
小鼠小鼠, 7天Cerebellar stem cellsPapain: 10u/mlDulbecco's PBSLee, A, Kessler, J, Read, T, Kaiser, C, Corbeil, D, Huttner ,W, Johnson, J, Wechsler-Reya, R.: Isolation of Neural Stem Cells from the Postnatal Cerebellum, Nat Neurosci 8, 723, 2005
Deoxyribonuclease I: 250 u/ml
小鼠小鼠, 胚胎Embryonic fibroblast feeder cellsCollagenase Type 4: 0.1%DMEMSchatten Gerald, Smith Joseph, Navara Christopher, Park Jong-Hyuk, Pedersen Roger:Culture of human embryonic stem cells, Nat Methods 2, 455-63, 2005
小鼠小鼠, 1天Neural progenitorPDS kit: per instructionssee referenceSeaberg, R., Smukler, S. and Van der Kooy, D.: Intrinsic Differences Distinguish Transiently Neurogenic Progenitors from Neural Stem Cells in the Early Postnatal Brain., Dev Biol 278, 71, 2005
小鼠小鼠, 8-10 周Liver derived stem cellsCollagenase Type 1: 0.1%HBSSKotton, D., Fabian, A. and Mulligan, R.: A Novel Stem-Cell Population in Adult Liver with Potent Hematopoietic-Reconstitution Activity., Blood 106, 1574, 2005
Neutral Protease: 2.4 u/ml
小鼠小鼠Bone marrowCollagenase Type 1: 0.15%PBSBertoncello, I. and Williams, B.: Hematopoietic Stem Cell Characterization by Hoechst 33342 and Rhodamine 123 Staining., Methods Mol Biol 263, 181, 2004
Neutral Protease: 0.15%
小鼠小鼠, 4-7 天Pluripotent stem cellsCollagenase Type 1: 220 u/mlMEMHowell, J., Lee, W., Morrison, P., Zhong, J., Yoder, M. and Srour, E.: Pluripotent Stem Cells Identified in Multiple Murine Tissues., Ann N Y Acad Sci 996, 158, 2003
Neutral Protease: 33 u/ml
小鼠小鼠, 15 天 或 3 個(gè)月Neural stem cellsPapain: 0.1%DMEM/F-12Gritti, A, Bonfanti, L, Doetsch, F, Caille, I, Alvarez-Buylla, A, Lim, D, Galli, R, Verdugo J, Herrera, D, and Vescovi A.: Multipotent Neural Stem Cells Reside into the Rostral Extension and Olfactory Bulb of Adult Rodents, J Neurosci 22(2), 437, 2002
小鼠小鼠, 胚胎Cortical progenitorsPDS kit: per instructionsSerum free mediumEstivill-Torrus, G., Pearson, H., Van Heyningen, V., Price, D. and Rashbass, P.: Pax6 is Required to Regulate the Cell Cycle and the Rate of Progression from Symmetrical to Asymmetrical Division in Mammalian Cortical Progenitors., Development 129, 455, 2002
小鼠小鼠, 4-8 個(gè)月Neural subventricular zoneTrypsin: 0.13%DMEM/F12Gritti A, Frolichsthal-Schoeller P, Galli R, Parati E, Cova L, Pagano S, Bjornson C, and Vescovi A.: Epidermal and Fibroblast Growth Factors Behave as Mitogenic Regulators for a Single Multipotent Stem Cell-like Population from the Subventricular Region of the Adult Mouse forebrain, J Neurosci 19(9), 3287, 1999
Hyaluronidase: 0.067%
小鼠小鼠, 6 周Muscle hematopoietic stem cellsCollagenase: 0.2%DMEMJackson, K., and Goodell, M.: Hematopoietic Potential of Stem Cells Isolated from Murine Skeletal Muscle, Proc Natl Acad Sci U S A 96, 14482, 1999
Trypsin: 0.1%
豬, 3-4 個(gè)月Synovial membrane and skin stem cellsCollagenase: 0.2%DMEMAndo, W., Kutcher, J., Krawetz, R., Sen, A., Nakamura, N., Frank, C. and Hart, D.: Clonal Analysis of Synovial Fluid Stem Cells to Characterize and Identify Stable Mesenchymal Stromal Cell/Mesenchymal Progenitor Cell Phenotypes in a Porcine Model: A Cell Source with Enhanced Commitment to the Chondrogenic Lineage., Cytotherapy 16, 776, 2014
豬,雌性, 小于一歲Adipose mesenchymal stemCollagenase Type 1: 0.1%DMEMWilliams, K., Picou, A., Kish, S., Giraldo, A., Godke, R. and Bondioli, K: Isolation and Characterization of Porcine Adipose Tissue-Derived Adult Stem Cells., Cells Tissues Organs188, 251, 2008
兔, 新西蘭, 8-10 周Tenocytes and tendon stem cellsCollagenase Type 1: 0.3%DMEMZhang, J. and Wang, J.: Characterization of Differential Properties of Rabbit Tendon Stem Cells and Tenocytes., BMC Musculoskelet Disord 11, 10, 2010
Neutral Protease: 0.4%
大鼠大鼠Mesenchymal Stem CellsCollagenase Type 1: 0.075%DMEMEmre, E, Yuksel, N, Duruksu, G, Pirhan, D, Subasi, C, Erman, G and Karaoz, E:Neuroprotective Effects of Intravitreally Transplanted Adipose Tissue and Bone Marrow-Derived Mesenchymal Stem Cells in an Experimental Ocular Hypertension Model.,Cytotherapy 17, 543-59, 2015
大鼠SD大鼠, 5 個(gè)月Mesenchymal Stromal CellsCollagenase Type 2: 0.075%DMEMVeronesi, F, Torricelli, P, Della, B, Pagani, S and Fini, M: In Vitro Mutual Interaction Between Tenocytes and Adipose-Derived Mesenchymal Stromal Cells., Cytotherapy 17, 215-23, 2015
大鼠大鼠Adipose Derived StemCollagenase Type 1: 0.1%DMEMWei, X., Du, Z., Zhao, L., Feng, D., Wei, G., He, Y., Tan, J., Lee, W., Hampel, H., Dodel, R., Johnstone, B., March, K., Farlow, M. and Du, Y.: IFATS Collection: The Conditioned Media of Adipose Stromal Cells Protect Against Hypoxia-Ischemia-Induced Brain Damage In Neonatal Rats., Stem Cells 27, 478, 2009
大鼠SD大鼠, 新生Neutral stem cells, Schwann cellsTrypsin: 0.25%DMEM/F12Zeng Yuan-Shan, Ding Ying, Wu Li-Zhi, Guo Jia-Song, Li Hai-Biao, Wong Wai-Man, Wu Wu-Tian: Co-transplantation of schwann cells promotes the survival and differentiation of neural stem cells transplanted into the injured spinal cord, Dev Neurosci 27, 20-6, 2005
Collagenase: 0.16%
大鼠大鼠, 雄性, 200-250 克Neural stem cellsPapain: 0.09%EBSSGobbel GT, Choi SJ, Beier S, Niranjan A: Long-term c*tion of multipotential neural stem cells from adult rat subependyma, Brain Res 980, 221, 2003
Deoxyribonuclease I: 0.1%
大鼠大鼠, 出生后的, 成年的Gut Neural Crest Stem CellsCollagenase Type 4: 0.1%HBSSKruger, G., Mosher, J., Bixby, S., Joseph, N., Iwashita, T. and Morrison, S.: Neural Crest Stem Cells Persist in the Adult Gut but Undergo Changes in Self-Renewal, Neuronal Subtype Potential, and Factor Responsiveness., Neuron 35, 657, 2002
Trypsin: 0.025%
大鼠大鼠, 成年的Neural stem cellsPapain: 2.5 u/mlDMEM/F-12Palmer, T., Markakis, E., Willhoite, A., Safar, F., and Gage, F.: Fibroblast Growth Factor-2 Activates a Latent Neurogenic Program in Neural Stem Cells from Diverse Regions of the Adult CNS, J Neurosci 19, 8487, 1999
Deoxyribonuclease I: 250 u/ml
Neutral Protease: 1 u/ml
大鼠大鼠Central nervous system stem cellsPapain: 0.01%DMEM/F-12Ray, J. and Gage, F.: Neural Stem Cell Isolation, Characterization and Transplantation,Modern Techniques in Neuroscience Research,Johansson, H. and Windhorst, U., Springer-Verlag New York, 339, 1999
Neutral Protease: 0.1%
大鼠大鼠, 胚胎Neurons and progenitorPapain: 20 u/mlEBSSMaric, D., Maric, I. and Barker, J.: Buoyant Density Gradient Fractionation and Flow Cytometric Analysis of Embryonic Rat Cortical Neurons and Progenitor Cells., Methods 16, 247, 1998

 

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6:我們還是Santa,Advanced Biotechnologies Inc, Worthington Biochemical ,bangs,BBInternational,crystalchem,dianova,FD Neurotechnologies,Inc. Dako,Inc, GePromegaridege, Glycotope Biotechnology GmbH; iduron,Innovative Research of America, Ludger, neuroprobe,Worthington Biochemical , Polysciences,prospecbi, QA-BIO,quickzyme,FULLERLABS,INC,sterlitech;sysy,TriLink BioTechnologies,Inc;worthington-biochem,zyagen等幾十家國(guó)外公司授權(quán)代理。
7:我們還是invitrogen,qiagen,Worthington Biochemical am,sigma;Promega,roche,merck, rnd,BD, GE,pierce,BioLegend等*批發(fā),歡迎合作。

 

 

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