Worthington 骨骼组织分离指南

发布者：艾美捷科技发布时间：2022-05-31

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原代细胞分离中，针对骨骼组织涉及到软骨细胞、成骨细胞、骨和骨髓基质细胞、股骨近端等等的解离通常会使用酶，如何选择合适的酶来完成对应骨骼组织的解离，以及原代细胞采集的常规操作， 艾美捷针对骨骼解离分种属、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：

本指南总结了Worthington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。

骨骼
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
Bovine	软骨细胞 / Chondrocytes	Collagenase Type 2: 0.4%	DMEM	Buschmann, M., Gluzband, Y., Grodzinsky, A., and Hunziker, E.: Mechanical Compression Modulates Matrix Biosynthesis in Chondrocyte/Agarose Culture, J Cell Sci 108, 1497, 1995
Chick, day old	成骨细胞 / Osteoblasts	Trypsin: 0.03%	DMEM	Gay, C., Lloyd, Q., and Gilman, V.: Characteristics and Culture of Osteoblasts Derived From Avian Long Bone, In Vitro Cell Dev Biol 30A, 379, 1994
Chick, Peterson/Arbor Acre, male, 4 weeks old (Gallus domesticus)	软骨细胞 / Chondrocytes	Trypsin: 0.25%	Ham's F12	Rosselot, G., Reginato, A.M., and Leach, R.M.: Development of a Serum-Free System to Study the Effect of Growth Hormone and Insulinlike Growth Factor-1 on Cultured Postembryonic Growth Plate Chondrocytes, In Vitro Cell Dev Biol 28A, 235, 1992
Chick, embryo	椎骨成软骨细胞 / Vertebrae chondroblasts	Trypsin: 0.25%	Simm's, CMF	Schiltz, J.R., Mayne R., and Holtzer, H.: The Synthesis of Collagen and Glycosaminoglycans by Dedifferentiated Chondroblasts in Culture, Differentiation 1, 97, 1973
Human	骨-软骨-基质细胞 / Bone-cartilage-stromal	Collagenase Type 2: 0.75%	HBSS	Agarwal, S., Loder, S., Sorkin, M., Li, S., Shrestha, S., Zhao, B., Mishina, Y., James, A. and Levi, B.: Analysis of Bone-Cartilage-Stromal Progenitor Populations in Trauma Induced and Genetic Models of Heterotopic Ossification., Stem Cells 34, 1692-701, 2016
Human	成骨细胞 / Osteoblasts	Collagenase Type 2: 0.2%； Trypsin: 0.25%-1.0%	DMEM	Taylor, S., Shah, M. and Orriss, I.: Generation of Rodent and Human Osteoblasts., Bonekey Rep 3, 585, 2014
Human	成骨细胞 / Osteoblasts	Collagenase Type 3: 0.2%	MEM	Kode, A.,Manavalan1, J.,Mosialou1, I., Bhagat G., Rathinam, C, Luo, N et al.: Leukemogenesis Induced by an Activating β-catenin Mutation in Osteoblasts, Nature 506, 240, 2014
Human, 22-73 year	成骨细胞 / Osteoblasts	Trypsin: 0.5%	Basal Medium	Kneser U, Voogd A, Ohnolz J, Buettner O, Stangenberg L, Zhang YH, Stark GB, Schaefer DJ: Fibrin gel-immobilized primary osteoblasts in calcium phosphate bone cement: in vivo evaluation with regard to application as injectable biological bone substitute, Cells Tissues Organs 179, 158-69, 2005
Human, 60+ year	骨细胞，成骨细胞 / Bone Cells, Osteoblasts	Collagenase Type 2: 200-250 u/ml	DMEM	Chen X., Qian H., Neff L., Satomura K., and Horowitz M.: Thy-1 antigen Expression by Cells in the Osteoblast Lineage, J Bone Miner Res 14, 362, 1999
Human	成骨细胞 / Osteoblasts	Trypsin: 0.1%	DMEM	Meikle, M., Boyd, S., Hembry, R., Compston, J., Croucher, P., and Reynolds, J.: Human Osteoblasts in Culture Synthesize Collagenase and Other Matrix Metalloproteinases in Response to Osteotropic Hormones and Cytokines, J Cell Sci 103, 1093, 1992
Human	成骨细胞 / Osteoblasts	Collagenase Type 4: 250 u/ml	DMEM	Fedarko, N.S., Termine, J.D., Young, M.F. and Robey, P.G.: Temporal Regulation of Hyaluronan and Proteoglycan Metabolism by Human Bone Cells in Vitro, J Biol Chem 265, 12200, 1990
Mouse	骨和骨髓基质细胞 / Bone and bone marrow stromal	Stemxyme 1: 0.1%； Neutral Protease: 0.1%	PBS	Mohrin, D., Koukos, G., Alabdulaaly, L., Paw, J., Hake, K., Kolumam, G., Bouxsein, M., Baron, R., Kutskova, Y., Freund, A., Liu, J., Zavala-Solorio, J., Bhargava, S., Maxwell, Trumble, J., Brito, A., Hu, D. and Brooks, D.: Inhibition of Longevity Regulator PAPP-A Modulates Tissue Homeostasis via Restraint of Mesenchymal Stromal Cells.,Aging Cell Vol. 20,, , e13313, 2021
Mouse	骨和骨髓基质细胞 / Bone and bone marrow stroma	Stemxyme 1: 0.1%； Neutral Protease: 0.1%	Media 199	Baryawno, N., Przybylski, D., Kowalczyk, M., Kfoury, Y., Severe, N., Gustafsson, K., Kokkaliaris, K., Mercier, F., Tabaka, M., Hofree, M., Dionne, D., Papazian, A., Lee, D., Ashenberg, O. and Subramanian, A.: A Cellular Taxonomy of the Bone Marrow Stroma in Homeostasis and Leukemia., Cell 177, 1915-1932.e16, 2019
Mouse	成骨细胞 / Osteoblast	Collagenase Type 1: 0.3%	HBSS	Wang, W., Majihail, G., Lui, C. and Zhou, L.: Osteoblast Sorting and Intracellular Staining of CXCL12., Bio Protoc 8, , 2018
Mouse	骨-软骨-基质细胞 / Bone-cartilage-stromal	Collagenase Type 2: 0.75%	HBSS	Agarwal, S., Loder, S., Sorkin, M., Li, S., Shrestha, S., Zhao, B., Mishina, Y., James, A. and Levi, B.: Analysis of Bone-Cartilage-Stromal Progenitor Populations in Trauma Induced and Genetic Models of Heterotopic Ossification., Stem Cells 34, 1692-701, 2016
Mouse	骨基质细胞 / Bone stromal	Collagenase Type 1: 0.3%	DMEM	Igarashi, H., Akahoshi, N., Ohto-Nakanishi, T., Yasuda, D. and Ishii, S.: The Lysophosphatidic Acid Receptor LPA4 Regulates Hematopoiesis-Supporting Activity of Bone Marrow Stromal Cells., Sci Rep 5, 11410, 2015
Mouse	成骨细胞 / Osteoblasts	Collagenase Type 1: 1.0%； Neutral Protease: 1.6%	MEM	Lee, D., Choung, H., Kim, H., Gronostajski, R., Yang, Y., Ryoo, H., Lee, Z., Kim, H., Cho, E. and Park, J.: NFI-C Regulates Osteoblast Differentiation Via Control of Osterix Expression., Stem Cells 32, 2467-79, 2014
Mouse, 2-5 day	成骨细胞 / Osteoblasts	Collagenase Type 2: 0.2%； Trypsin: 0.25%-1.0%	DMEM	Taylor, S., Shah, M. and Orriss, I.: Generation of Rodent and Human Osteoblasts., Bonekey Rep 3, 585, 2014
Mouse	成骨细胞 / Osteoblasts	Collagenase Type 3: 0.2%	MEM	Kode, A.,Manavalan1, J.,Mosialou1, I., Bhagat G., Rathinam, C, Luo, N et al.: Leukemogenesis Induced by an Activating β-catenin Mutation in Osteoblasts, Nature 506, 240, 2014
Mouse, 4-22 month	骨细胞 / Osteocytes	Collagenase Type 1: 300 u/ml	MEM	Stern, A., Stern, M., Van Dyke, M., Jahn, K., Prideaux, M. and Bonewald, L.: Isolation and Culture of Primary Osteocytes from the Long Bones of Skeletally Mature and Aged Mice., Biotechniques 52, 361, 2012
Mouse	骨内膜细胞 / Endosteal cells	Collagenase Type 1: 0.3%	DMEM	Nakamura, Y., Arai, F., Iwasaki, H., Hosokawa, K., Kobayashi, I., Gomei, Y., Matsumoto, Y., Yoshihara, H. and Suda, T.: Isolation and Characterization of Endosteal Niche Cell Populations that Regulate Hematopoietic Stem Cells., Blood 116, 1422, 2010
Mouse	骨髓 / Bone marrow	Collagenase: 0.2%	DMEM	Morikawa, S., Mabuchi, Y., Kubota, Y., Nagai, Y., Niibe, K., Hiratsu, E., Suzuki, S., Miyauchi-Hara, C., Nagoshi, N., Sunabori, T., Shimmura, S., Miyawaki, A., Nakagawa, T., Suda, T., Okano, H. and Matsuzaki, Y.: Prospective Identification, Isolation, and Systemic Transplantation of Multipotent Mesenchymal Stem Cells in Murine Bone Marrow., J Exp Med 206, 2483-96, 2009
Mouse, male, 6-8 week old	破骨细胞 / Osteoclasts	Collagenase Type 3: 0.1%	DMEM	Sakai, E., Miyamoto, H., Okamoto, K., Kato, Y., Yamamoto, K., and Sakai, H.: Characterization of Phagosomal Subpopulations Along Endocytic Routes in Osteoclasts and Macrophages, J. Biochem. 130, 823, 2001
Mouse, 3-5 day, 6-8 week	骨细胞，成骨细胞 / Bone Cells, Osteoblasts	Collagenase Type 2: 200-250 u/ml	DMEM	Chen X., Qian H., Neff L., Satomura K., and Horowitz M.: Thy-1 antigen Expression by Cells in the Osteoblast Lineage, J Bone Miner Res 14, 362, 1999
Mouse, BALB/c	成骨细胞样细胞，基质细胞系，造血母细胞 / Osteoblast-like Cells，Stromal Cell Lines，Hematopoietic Blast Cells	Trypsin: 0.1%	Eagle's MEM	Takanashi, H., Matsuishi, T., and Yoshizato, K.: Establishment and Characterization of Stromal Cell Lines That Support Differentiation of Murine Hematopoietic Blast Cells into Osteoblast-like Cells, In Vitro Cell Dev Biol 30A, 384, 1994
Mouse, Swiss-Webster	新生儿骨 / Neonatal bone	Collagenase Type 2: 0.20%	Tris-buffered saline	Chen,T. and Feldman,D.: Regulation of 1,25-Dihydroxyvitamin D3 Receptors in Cultured Mouse Bone Cells, J Biol Chem 256, 5561, 1981
Rat, 2 day	成骨细胞 / Osteoblasts	Collagenase Type 2: 0.1%； Trypsin: 0.25%	DMEM	Jeon, J., Lee, M. and Yang, H.: Differentiated Osteoblasts Derived Decellularized Extracellular Matrix to Promote Osteogenic Differentiation., Biomater Res 22, 4, 2018
Rat, Wistar, 250g	股骨近端 / Proximal femur stem	Collagenase Type 1: 0.075%	HBSS	Jacobs, F., Gijsen, H., Van de Vyver, M. and Ferris, W.: Vanadate Impedes Adipogenesis in Mesenchymal Stem Cells Derived from Different Depots within Bone., Front Endocrinol 7, 108, 2016
Rat, 2-3 day	成骨细胞 / Osteoblasts	Collagenase Type 2: 0.2%； Trypsin: 0.25%-1.0%	DMEM	Taylor, S., Shah, M. and Orriss, I.: Generation of Rodent and Human Osteoblasts., Bonekey Rep 3, 585, 2014
Rat, fetal, 21 days of gestation	卡尔瓦利亚 / Calvaria	Collagenase: 0.2%	MEM	Owen, T., Aronow, M., Shalhoub, V., Barone, L., Wilming, L., Tassinari, M., Kennedy, M., Pockwinse, S., Lian, J., and Stein, G.: Progressive Development of the Rat Oseoblast Phenotype In Vitro: Reciprocal Relationships in Expression of Genes Associated with Osteoblast Proliferation and Differentiation During Formation of the Bone Extracellular Matrix, J Cell Physiol 143, 420, 1990
Rat, newborn	成骨细胞样细胞 / Osteoblastlike cells	Collagenase Type 2: 0.3%	MEM	Ernst, M., and Froesch, E.: Osteoblastlike Cells in a Serum-Free Methylcellulose Medium Form Colonies: Effects of Insulin and Insulinlike Growth Factor I, Calcif Tissue Int 40, 27, 1987
Rat, fetus, 17-21 day	卡尔瓦利亚 / Calvaria	Collagenase: 0.01%-0.6%	Tris-buffered saline	Peck, W., Birge, S., and Fedak, S.: Bone Cells: Biochemical and Biological Studies After Enzymatic Isolation, Science 146, 1476, 1964