Worthington 骨骼組織分離指南

發(fā)布者：艾美捷科技發(fā)布時(shí)間：2022-05-31

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原代細(xì)胞分離中，針對骨骼組織涉及到軟骨細(xì)胞、成骨細(xì)胞、骨和骨髓基質(zhì)細(xì)胞、股骨近端等等的解離通常會(huì)使用酶，如何選擇合適的酶來完成對應(yīng)骨骼組織的解離，以及原代細(xì)胞采集的常規(guī)操作， 艾美捷針對骨骼解離分種屬、不同的細(xì)胞、酶組合推薦，推薦濃度、培養(yǎng)基以及參考文獻(xiàn)，詳細(xì)匯總指南：

本指南總結(jié)了Worthington本技術(shù)指南描述了標(biāo)準(zhǔn)實(shí)驗(yàn)室程序；為建立細(xì)胞分離方案提供了一種合乎邏輯的實(shí)驗(yàn)方法；并列出了許多組織特異性參考資料，以幫助開發(fā)有效的方法。更多其他組織的解離指南，點(diǎn)擊查看：各組織分離指南。

骨骼
種屬詳情	細(xì)胞名稱	酶組合推薦，含推薦濃度	培養(yǎng)基	參考文獻(xiàn)
Bovine	軟骨細(xì)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	成骨細(xì)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)	軟骨細(xì)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	椎骨成軟骨細(xì)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-軟骨-基質(zhì)細(xì)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	成骨細(xì)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	成骨細(xì)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	成骨細(xì)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	骨細(xì)胞，成骨細(xì)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	成骨細(xì)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	成骨細(xì)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	骨和骨髓基質(zhì)細(xì)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	骨和骨髓基質(zhì)細(xì)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	成骨細(xì)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-軟骨-基質(zhì)細(xì)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	骨基質(zhì)細(xì)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	成骨細(xì)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	成骨細(xì)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	成骨細(xì)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	骨細(xì)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	骨內(nèi)膜細(xì)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	破骨細(xì)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	骨細(xì)胞，成骨細(xì)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	成骨細(xì)胞樣細(xì)胞，基質(zhì)細(xì)胞系，造血母細(xì)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	新生兒骨 / 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	成骨細(xì)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	成骨細(xì)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	卡爾瓦利亞 / 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	成骨細(xì)胞樣細(xì)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	卡爾瓦利亞 / 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