原代细胞分离中，针对其他组织涉及到髓核，肌腱细胞、牙髓干细胞、口腔上皮等等的解离通常会使用酶，如何选择合适的酶来完成对应其他组织的解离，以及原代细胞采集的常规操作， 艾美捷针对其他相关的组织分类解离分种属、不同的细胞、酶组合推荐，推荐浓度、培养基以及参考文献，详细汇总指南：
本技术指南描述了标准实验室程序； 为建立细胞分离方案提供了一种合乎逻辑的实验方法； 并列出了许多组织特异性参考资料，以帮助开发有效的方法。更多其他组织的解离指南，点击查看：各组织分离指南。
| 其他组织 | |||||
| 种属 | 种属详情 | 细胞名称 | 酶组合推荐，含推荐浓度 | 培养基 | 参考文献 |
| 牛 | Bovine | 髓核 / Nucleus pulposus | Collagenase Type 2: 0.2% | DMEM | Lee, J., Cheung, K. and Leung, V.: Systematic Study of Cell Isolation from Bovine Nucleus Pulposus: Improving Cell Yield and Experiment Reliability., J Orthop Res 33, 1743-55, 2015 |
| 马 | Equine | 肌腱细胞 / Tendon Cells | Collagenase Type 1: 0.1% | DMEM | Nemoto, M., Kizaki, K., Yamamoto, Y., Oonuma, T. and Hashizume, K.: Tenascin-C Expression in Equine Tendon-derived Cells During Proliferation and Migration., J Equine Sci 24, 17-24, 2013 |
| 山羊 | Goat | 髌下脂肪垫间充质基质细胞 / Infrapatellar fat pad mesenchymal stromal | Collagenase: 0.15% | DMEM | Arora, A., Sriram, M., Kothari, A. and Katti, D.: Co-culture of Infrapatellar Fat Pad-Derived Mesenchymal Stromal Cells and Articular Chondrocytes in Plasma Clot for Cartilage Tissue Engineering., Cytotherapy 19, 881-894, 2017 |
| 人 | Human | 粘附成纤维细胞 / Adhesion fibroblasts | Collagenase Type 4: 0.2% | Medium 199 | Foster, D., Marshall, C., Gulati, G., Chinta, M., Nguyen, A., Salhotra, A., Jones, R., Burcham, A., Lerbs, T., Cui, L., King, M., Titan, A., Ransom, R., Manjunath, A., Hu, M. and Blackshear, C.: Elucidating the Fundamental Fibrotic Processes Driving Abdominal Adhesion Formation., Nat Commun 11, 4061, 2020 |
| 人 | Human | 髓核 / Nucleus pulposus | Pronase: 0.25%； Collagenase Type 2: 600 u/ml |
DMEM | Sun, Y., Lv, M., Zhou, L., Tam, V., Lv, F., Chan, D., Wang, H., Zheng, Z., Cheung, K. and Leung, V.: Enrichment of Committed Human Nucleus Pulposus Cells Expressing Chondroitin Sulfate Proteoglycans under Alginate Encapsulation., Osteoarthritis Cartilage 23, 1194-203, 2015 |
| 人 | Human, 3-7 year | 牙髓干细胞 / Dental pulp stem cells | Collagenase Type 1: 0.3%； Neutral Protease: 0.4% |
DMEM | Salmon, B., Bardet, C, Khaddam, M. Naji, J., Coyac, B., Baroukh, B., Letourneur, F., Lesieur, J., Decup, F., Le Denmat, D., Nicoletti, A., Poliard, A, Rowe, P., Huet, E., Vital, S. and Linglart, A.: MEPE-Derived ASARM Peptide Inhibits Odontogenic Differentiation of Dental Pulp Stem Cells and Impairs Mineralization in Tooth Models of X-linked Hypophosphatemia., PLoS ONE 8, e56749, 2013 |
| 人 | Human | 牙髓干细胞 / Dental pulp stem | Collagenase Type 1: 0.3%； Neutral Protease: 0.4% |
DMEM/F12 | Bonnamain, 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 |
| 人 | Human | 间充质基质细胞 / Mesenchymal stromal | Collagenase Type 1: 0.075% | PBS | Lee, Y., Lee, J., Park, H., Lim, Y., Lee, J., Wang, S. and Lee, B.: Isolation of Mesenchymal Stromal Cells (MSCs) from Human Adenoid Tissue., Cell Physiol Biochem 31, 513-24, 2013 |
| 人 | Human | 菌状味乳头 / Fungiform taste papillae | Collagenase Type 1: 550 u/ml； Elastase: 10 u/ml； Soybean Trypsin Inhibitor: 0.09% |
see reference | Ozdener, H., Spielman, A. and Rawson, N.: Isolation and Culture of Human Fungiform Taste Papillae Cells, J Vis Exp 63, e3730, 2012 |
| 人 | Human | 肌腱细胞 / Tendon cells | Collagenase Type 2: 0,3% | DMEM/Hams F12 | Pauly, S., Klatte, F., Strobel, C., Schmidmaier, G., Greiner, S. and Scheibel, M. Wildemann B: Characterization of Tendon Cell Cultures of the Human Rotator Cuff., Eur Cell Mater 20, 84-97, 2010 |
| 人 | Human | 脂肪来源的基质细胞 / Adipose derived stromal | Collagenase Type 2: 0.075% | DMEM | Panetta, N., Gupta, D., Kwan, M., Wan, D., Commons, G. and Longaker, M.: Tissue Harvest by Means of Suction-Assisted or Third-Generation Ultrasound-Assisted Lipoaspiration Has No Effect on Osteogenic Potential of Human Adipose-Derived Stromal Cells., Plast Reconstr Surg 124, 65, 2009 |
| 人 | Human | 口腔上皮 / Oral epithelial | Neutral Protease: 1.2 u/ml； Trypsin: 0.05% |
PBS | Nakamura, T., Endo, K. and Kinoshita, S.: Identification of Human Oral Keratinocyte Stem/Progenitor Cells by Neurotrophin Receptor p75 and the Role of Neurotrophin/p75 Signaling., Stem Cells 25, 628-38, 2007 |
| 人 | Human | 颈动脉斑块巨噬细胞 / Carotid artery plaque macrophage | Collagenase Type 4: 450 u/ml； Deoxyribonuclease I: 500 u/ml； Soybean Trypsin Inhibitor: 0.1% |
HBSS | Patino WillmarD, Kang Ju-Gyeong, Matoba Satoaki, Mian OmarY, Gochuico BernadetteR, Hwang PaulM: Atherosclerotic plaque macrophage transcriptional regulators are expressed in blood and modulated by tristetraprolin, Circ Res 98, 1282-9, 2006 |
| 人 | Human | 食管微血管内皮 / Esophageal microvascular endothelial | Collagenase Type 2: 0.2% | MCDB-131 | Rafiee, P., Ogawa, H., Heidemann, J., Li, M., Aslam, M., Lamirand, T., Fisher, P., Graewin, S., Dwinell, M., Johnson, C., Shaker, R. and Binion, D.: Isolation and Characterization of Human Esophageal Microvascular Endothelial Cells: Mechanisms of Inflammatory Activation., Am J Physiol Gastrointest Liver Physiol 285, G1277, 2003 |
| 人 | 胃上皮细胞 / Gastric epithelial cells | Collagenase Type 2: 200 u/ml； Neutral Protease: 1.2 u/ml； Soybean Trypsin Inhibitor: 0.125% |
L-15 | Smoot, D., Sewchand, J., Young, K., Desbrodes, B., Allen, C. and Naab, T.: A Method for Establishing Primary Cultures of Human Gastric Epithelial Cells, Meth Cell Sci 22, 133, 2000 | |
| 人 | Human | 滑膜细胞 / Synoviocytes | Collagenase: 0.2% | DMEM/F12 | Chen V, Croft D, Purkis P, Kramer IM: Co-culture of synovial fibroblasts and differentiated U937 cells is sufficient for high interleukin-6 but not interleukin-1beta or tumour necrosis factor-alpha release., Br J Rheumatol 37, 148-56, 1998 |
| 人 | Human | 消化细胞 / Peptic cells | Collagenase Type 4: 0.1%； Soybean Trypsin Inhibitor: 0.2% |
Ringer solution | Lanas. A., Nerin, J., Esteva, F. and Sainz, R.: Non-Steroidal Anti-Inflammatory Drugs and Prostaglandin Effects on Pepsinogen Secretion by Dispersed Human Peptic Cells., Gut 36, 657-63, 1995 |
| 人 | Human | 根尖肉芽肿 / Periapical granuloma | CLSPA: 0.25% | RPMI-1640 | Stern MH, Dreizen S, Mackler BF, Levy BM: Isolation and characterization of inflammatory cells from the human periapical granuloma, J Dent Res 61, 1408-12, 1982 |
| 昆虫 | Insect, Lepidoptera | 鳞翅目 / Lepitopteran | Collagenase Type 3: 0.35%； Hyaluronidase: 0.01% |
Dulbecco PBS | Goodwin, R and McCawley, P: Initiating Attached Cell Lines From the Lepidoptera (Insecta), Meth Cell Sci 3, 567, 1977 |
| 无脊椎动物 | Squid | 地幔 / Mantle | Collagenase: see reference； Papain: see reference |
Phosphate buffer | Raman, M. and Mathew, S.: Study of Chemical Properties and Evaluation of Collagen in Mantle, Epidermal Connective Tissue and Tentacle of Indian Squid, Loligo duvauceli Orbigny., J Food Sci Technol 51, 1509-16, 2014 |
| 小鼠 | Mouse | 类器官培养物 / Organoid cultures | Stemxyme 1: 0.1% | PBS | Huang, L., Desai, R., Conrad, D., Leite, N., Akshinthala, D., Lim, C., Gonzalez, R., Muthuswamy, L., Gartner, Z. and Muthuswamy, S.: Commitment and Oncogene-induced Plasticity of Human Stem Cell-derived Pancreatic Acinar and Ductal Organoids.,Cell Stem Cell Vol. 28,, , 1090-1104.e6, 2021 |
| 小鼠 | Mouse, embryonic | 气管 / Trachea | Elastase: 0.1%； Neutral Protease: 0.25%； Deoxyribonuclease I: 0.01%； Trypsin: 0.125% |
DMEM/F12 | He, M., Wu, B., Ye, W., Le, D., Sinclair, A., Padovano, V., Chen, Y., Li, K., Sit, R., Tan, M., Caplan, M., Neff, N., Jan, Y., Darmanis, S. and Jan, L.: Chloride Channels Regulate Differentiation and Barrier Functions of the Mammalian Airway., Elife 9, 53085, 2020 |
| 小鼠 | Mouse | 粘附成纤维细胞 / Adhesion fibroblasts | Collagenase Type 4: 0.2% | Medium 199 | Foster, D., Marshall, C., Gulati, G., Chinta, M., Nguyen, A., Salhotra, A., Jones, R., Burcham, A., Lerbs, T., Cui, L., King, M., Titan, A., Ransom, R., Manjunath, A., Hu, M. and Blackshear, C.: Elucidating the Fundamental Fibrotic Processes Driving Abdominal Adhesion Formation., Nat Commun 11, 4061, 2020 |
| 小鼠 | Mouse, 8-12 week | 膀胱 / Urinary bladder | Collagenase Type 6: 20 u/mlCollagenase Type 1: 50 u/ml； Collagenase Type 3: 40 u/ml； Papain: 45 u/ml； Deoxyribonuclease I: 4 u/ml； |
Tyrode solution | Atta, H., Gheinani, A., Wacker, A., Heshmati, Y., Bigger-Allen, A., Lambrinos, G., Gao, Y., Bielenberg, D. and Adam, R.: A Single Cell Dissociation Approach for Molecular Analysis of Urinary Bladder in the Mouse Following Spinal Cord Injury., J Vis Exp , , 2020 |
| 小鼠 | Mouse | 尿路上皮 / Urothelial | Collagenase Type 1: 0.5% | PBS | Huang, C., Chen, C. and Shyr, C.: The Anti-Tumor Effect of Intravesical Administration of Normal Urothelial Cells on Bladder Cancer., Cytotherapy 19, 1233-1245, 2017 |
| 小鼠 | Mouse | 骨髓基质细胞 / Bone marrow stromal | Collagenase Type 1: 0.2% | PBS | Ishida, T., Suzuki, S., Lai, C., Yamazaki, S., Kakuta, S., Iwakura, Y., Nojima, M., Takeuchi, Y., Higashihara, M., Nakauchi, H. and Otsu, M.: Pre-Transplantation Blockade of TNF-α-Mediated Oxygen Species Accumulation Protects Hematopoietic Stem Cells., Stem Cells 35, 989-1002, 2017 |
| 小鼠 | Mouse | 耳蜗 / Cochlear | Papain: 20 u/ml； Trypsin: 0.05% |
DMEM | Kim, Y., Wang, S., Tymanskyj, S., Ma, L., Tao, H. and Zhang, L.: Dcc Mediates Functional Assembly of Peripheral Auditory Circuits., Sci Rep 6, 23799, 2016 |
| 小鼠 | Mouse | 上皮干细胞 / Epithelial stem | Collagenase: 2% | DMEM/F12 | Chavez, M., Hu, J., Seidel, K., Li, C., Jheon, A., Naveau, A., Horst, O. and Klein, O.: Isolation and Culture of Dental Epithelial Stem Cells from the Adult Mouse Incisor., J Vis Exp , , 2014 |
| 小鼠 | Mouse, 26-30 g | 肌间丛 / Myenteric plexus | Collagenase Type 2: 0.13% | Neurobasal A | Smith, T., Grider, J., Dewey, W. and Akbarali, H.: Morphine Decreases Enteric Neuron Excitability via Inhibition of Sodium Channels., PLoS ONE 7, e45251, 2012 |
| 小鼠 | Mouse, 10 week | 滑膜间充质 / Synovial mesenchymal | Collagenase: 0.1%； Deoxyribonuclease I: 0.005% |
DMEM | Futami, 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 ONE 7, e45517, 2012 |
| 小鼠 | Mouse | 脾脏，骨髓内皮 / Spleen, bone marrow endothelial | Collagenase Type 4: 0.3-1.0%； Deoxyribonuclease I: 20 u/ml |
PBS | Shi, 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 |
| 小鼠 | Mouse | 树突状 / Dentritic | Collagenase Type 2: 0.1%； Deoxyribonuclease I: 0.001% |
DME | Stock, A., Booth, S. and Cerundolo, V.: Prostaglandin E2 Suppresses the Differentiation of Retinoic Acid-Producing Dendritic Cells in Mice and Humans., J Exp Med 208, 761, 2011 |
| 小鼠 | Mouse | 耳蜗 / Cochleaer | Trypsin: 0.125% | DMEM/F-12 | Jan, T., Chai, R., Sayyid, Z. and Cheng, A.: Isolating LacZ-Expressing Cells from Mouse Inner Ear Tissues Using Flow Cytometry., J Vis Exp 58, e3432, 2011 |
| 小鼠 | Mouse, 6-8 week | 骨髓间充质干细胞 / Bone marrow mesenchymal stem | Collagenase Type 1: 0.25% | RPMI 1640 | Xu, 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 | Mouse, 3-6 week | 耳间充质干细胞 / Ear mesenchymal stem | Collagenase Type 1: 0.2% | DMEM/F12 | Staszkiewicz, 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 | Mouse, 6-10 week | 唾液腺和胃 / Salivary gland and stomach | Collagenase Type 4: 0.8%； Deoxyribonuclease I: 1.0% |
RPMI-1640 | Ji, H., Rintelen, F., Waltzinger, C., Meier, D, Bilancio, A., Pearce, W., Hirsch, E., Wymann, M., Ruckle, T., Camps, M., Vanhaesebroeck, B., Okkenhaug, K. and Rommel, C.: Inactivation of PI3Kgamma and PI3Kdelta Distorts T-cell Development and Causes Multiple Organ Inflammation., Blood 110, 2940, 2007 |
| 小鼠 | Mouse | 骨髓 / Bone marrow | Collagenase Type 1: 0.15%； Neutral Protease: 0.15% |
PBS | Bertoncello, I. and Williams, B.: Hematopoietic Stem Cell Characterization by Hoechst 33342 and Rhodamine 123 Staining., Methods Mol Biol 263, 181, 2004 |
| 小鼠 | Mouse, 25-30 g | 气管炎症细胞 / Tracheal inflammatory cells | Collagenase Type 4: 0.1%； Deoxyribonuclease I: 50 u/ml； Soybean Trypsin Inhibitor: 0.1% |
RPMI 1640 | Minamoto Kanji, Pinsky DavidJ: Recipient iNOS but not eNOS deficiency reduces luminal narrowing in tracheal allografts, J Exp Med 196, 1321-33, 2002 |
| 猪 | Porcine, 6 month | 滑膜细胞 / Synoviocytes | Hyaluronidase: 660 u/ml； Trypsin: 0.25%； Collagenase Type 2: 583 u/ml |
DMEM | Kean, T and Dennis, J.: Synoviocyte Derived-Extracellular Matrix Enhances Human Articular Chondrocyte Proliferation and Maintains Re-Differentiation Capacity at Both Low and Atmospheric Oxygen Tensions., PLoS ONE 10, e0129961, 2015 |
| 猪 | Porcine, <10 week | 髓核 / Nucleus pulposus | Collagenase: 0.05%； Pronase: 0.3% |
DMEM | Potier, E. and Ito. K.: Can Notochordal Cells Promote Bone Marrow Stromal Cell Potential for Nucleus Pulposus Enrichment? A Simplified in Vitro System., Tissue Eng Part A 20, 3241-51, 2014 |
| 猪 | Porcine, various ages | 髓核和房颤 / Nucleous pulposus and AF | Pronase: 0.05%； Collagenase: 0.2% |
Modified F-12K | Cho, H., Park, S., Lee, S., Kang, M., Hasty, K. and Kim, S.: Snapshot of Degenerative Aging of Porcine Intervertebral Disc: A Model to Unravel the Molecular Mechanisms., Exp Mol Med 43, 334-40, 2011 |
| 兔 | Rabbit, New Zealand, 8-10 week | 肌腱细胞和肌腱干细胞 / Tenocytes and tendon stem cells | Collagenase Type 1: 0.3%； Neutral Protease: 0.4% |
DMEM | Zhang, J. and Wang, J.: Characterization of Differential Properties of Rabbit Tendon Stem Cells and Tenocytes., BMC Musculoskelet Disord 11, 10, 2010 |
| 大鼠 | Rat, Lewis, M | 滑膜巨噬细胞 / Synovial macrophages | Collagenase Type 1: 250 u/ml | DMEM | Jain, S., Tran, T. and Amiji, M.: Macrophage Repolarization with Targeted Alginate Nanoparticles Containing IL-10 Plasmid DNA for the Treatment of Experimental Arthritis., Biomaterials 61, 162-77, 2015 |
| 大鼠 | Rat, SD, 6-8 month | 多能肌腱 / Pluripotent tendon | Collagenase Type 1: 0.3%； Neutral Protease: 0.4% |
DMEM | Zhou, B., Zhou, Y. and Tang, K.: The Effects of Substance P on Pluripotent Tendon Cells: an In Vitro and In Vivo Study., J Musculoskelet Neuronal Interact 14, 349-58, 2014 |
| 大鼠 | Rat, Wistar, 7-9 week | 牙龈线粒体 / Gingival mitochondria | Collagenase Type 1: 0.115-0.130 % | HBSS | Kaneko, N., Rikimaru, T., Fujimura, T., Mori, S., Hidaka, S. and Kaya, H.: Preparation of Rat Gingival Mitochondria with an Improved Isolation Method., Int J Dent 2010, 275103, 2010 |
| 大鼠 | Rat, male, 250-350g | 颌下腺泡 / Submandibular acinar | Collagenase: 0.05% | PBS | Silver, N., Proctor, GB, Arno, M. and Carpenter GH: Activation of mTOR Coincides with Autophagy During Ligation-induced Atrophy in the Rat Submandibular Gland., Cell Death Dis 1, e14, 2010 |
| 大鼠 | Rat, SD, 8-10 week | 口腔成纤维细胞 / Oral fibroblasts | Collagenase Type 1: 200 u/ml | DMEM | Lin, A., Hokugo, A., Choi, J. and Nishimura, I.: Small Cytoskeleton-Associated Molecule, Fibroblast Growth Factor Receptor 1 Oncogene Partner 2/Wound Inducible Transcript-3.0 (FGFR1OP2/wit3.0), Facilitates Fibroblast-Driven Wound Closure., Am J Pathol 176, 108-21, 2010 |
| 大鼠 | Rat, 8-10 week | 滑膜细胞 / Synovial cells | Collagenase Type 1: 250 u/ml | RPMI 1640 | Moghaddami, M., Cleland, L. and Mayrhofer, G.: MHC II+ CD45+ Cells from Synovium-Rich Tissues of Normal Rats: Phenotype, Comparison with Macrophage and Dendritic Cell Lineages and Differentiation into Mature Dendritic Cells In Vitro., Int Immunol 17, 1103, 2005 |
