Primary cells of human small intestinal mucosal epithelium

Primary cells of human small intestinal mucosal epithelium

Training Information:

Wrapped condition: collagen I in mouse tail（2-5Mg/cm2）

culture mediumcontainFBSGrowth additivesPenicillinTheStreptomycinwait

Fluid change frequency per2-3Change the fluid once every day

Growth characteristics: Wall adhesion

Cell morphology epithelial cell like

Passage characteristics can be passed down2-3generation

digestive juice0.25%

Cultivation conditions gas phase: air,95%；CO2，5%

Isolation of human small intestinal mucosal epithelial cells from small intestinal tissue; The small intestine is located in the abdomen, with its upper end connected to the pylorus and the stomach, and its lower end connected to the large intestine through the trapdoor. It is the main site for food digestion and absorption. The small intestine is coiled in the abdominal cavity, connected to the pylorus and cecum, and divided into three parts: duodenum, jejunum, and ileum. Digestion in the small intestine is crucial because food undergoes chemical digestion of pancreatic juice, bile, and small intestinal fluid, as well as mechanical digestion of small intestinal motility, completing the digestion process and allowing nutrients to be absorbed by the small intestinal mucosa. The wall of the small intestine is composed of mucosa, submucosa, muscle layer, and serosa. Its structural characteristics are that the tube wall has circular folds, the mucosa has many villi, and the epithelium at the base of the villi sinks to the lamina propria, forming tubular intestinal glands with openings located between the villi roots. The villi and intestinal glands are closely related to the digestion and absorption functions of the small intestine; The cells that make up the intestinal gland include columnar cells, goblet cells, Paneth cells, and undifferentiated cells. Columnar cells and endocrine cells are similar to the villous epithelium, while columnar cells close to the villi are similar to absorptive cells. Columnar cells in the deep part of the villi have fewer and shorter microvilli, and do not form striated edges. The small intestine has three functions: digestion, absorption, secretion, and motility, with absorption and secretion being the main functions. The contraction of smooth muscle cells is the driving force behind intestinal peristalsis, promoting downward movement of food. Intestinal mucosal epithelial cells are an important barrier for the internal and external environment of the body, continuously exposed to a large amount of antigens, and also the first line of defense for the body against pathogenic microorganisms. Therefore, in addition to important physiological functions such as absorption, secretion, and transport, intestinal mucosal epithelial cells also play an important role in the innate and acquired immune defense mechanisms of the mucosa. As the first cells to come into contact with antigens, intestinal mucosal epithelial cells play a crucial role in the initial stage of mucosal immune response, determining the occurrence, nature, and intensity of mucosal immune response.

Method Introduction:

The human small intestinal mucosal epithelial cells isolated in the company's laboratory were prepared by mechanical separation followed by collagenase digestion, and screened through epithelial cell specific culture medium. The total number of cells was approximately5×10?cells/Bottle.

Quality inspection:

The human small intestine mucosal epithelial cells isolated from the company laboratoryCytokeratin-18Immunofluorescence identification, purity can reach90%Above, and not containingHIV-1TheHBVTheHCVMycoplasma, bacteria, yeast, fungi, etc.

① Organizational block cultivation method

Tissue block culture is a commonly used, simple, and highly successful primary culture method. The basic method is to inoculate the cut small tissue clusters into a culture bottle (or dish), and the bottle wall can be pre coated with a thin layer of collagen to facilitate the adhesion of the tissue blocks to the bottle wall, allowing surrounding cells to grow outward along the bottle wall.

② Digestive cultivation method

③ Suspension cell culture method

For cells that grow in suspension, such as leukemia cells, lymphocytes, bone marrow cells, cancer cells and immune cells in pleural and ascites, digestion is not required. They can be isolated by low-speed centrifugation and cultured directly, or cultured by inoculating after lymphocyte stratification.

④ Organ culture

Organ culture refers to the direct cultivation of organs or tissue blocks obtained from donors under specific environmental conditions outside the body without tissue separation. Organ culture can maintain the relative integrity of organ tissues and can be used to focus on observing the connections, arrangements, and interactions between cells, as well as the biological regulatory effects of local environments.

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PACAP(1-38 Peptide 0.5mgPACAP(1-38 Peptide)Adenylate cyclase activating peptide1-38(Whole protein)

ITGA5 & ITGB6 Protein HumanRecombinant humanITGA5 & ITGB6 Heterodimerprotein

HARecombinant influenza AH1N1 (A/New York/1/1918)hemagglutininHA1 (Hemagglutinin)proteinProtein

ACVR2B Protein HumanRecombinant humanACVR2B / ActivinR-IIBprotein(Fclabel)

CD97Recombinant humanCD97protein(Fclabel) Protein

人抑制素B(INH-B)ELISAtest kit

Human ansforming growth factor beta 2 (TGF beta 2) ELISA KitHuman transforming growth factor beta2(TGFB2)test kit

HumanDickkopf1,DKK1ELISAKitpersonDickheadf1(DKK1)test kit96T/48TImport packaging

HumanIerleukin11,IL-11Reagent kit for human interleukin11(IL-11)Kit specifications:96T/48T

Organizing the Kosaki virusA(CoxsackievirusA)quantitativePCRAmplification kit20time

HumanIschemiaModifiedAlbumin,IMAELISAKitHuman ischemia modified albuminIMAKit specifications:96T/48T

Human mucin2(MUC2)Enzyme linked immunosorbent assay kit Human MUC2 (Mucin 2) ELISA Kit

Human mucin20(MUC20)Enzyme linked immunosorbent assay kit Human MUC20 (Mucin 20) ELISA Kit

Human brain hormone receptor(C1)Enzyme linked immunosorbent assay kit Human C1 (Cannabinoid Receptor 1, Brain) ELISA Kit

Human adenylate cyclase related protein1(CAP1)Enzyme linked immunosorbent assay kit Human CAP1 (Adenylate Cyclase Associated Protein 1) ELISA Kit

Primary cells of human small intestinal mucosal epitheliumGuinea pig interleukin1alpha(IL1alpha)Reagent kit, English name:IL1alphaELISA Kit

Human mannose (Mannose) ELISA KitHuman mannose(Mannose)test kit

rabbitHyaluronicacid,HAELISAKitRabbit hyaluronic acid(HA)test kit96T/48TImport packaging

CLIAKitforATMA/TMAB(HumanAi-ThyroidMicrosomeAibody)ELISAKITHuman anti thyroid microsomal antibody

Mitochondrial complexIIprotein expressionELISAQuantitative reagent kit25time

Rabbitai-braiissueaibody,ABAbELISAKitRabbit anti brain tissue antibody(ABAb)Kit specifications:96T/48T

GP140Recombinant human immunodeficiency virus type IHIV-1 gp140protein(group M, subtype A, strain 92UG037.1) (Fclabel) Protein

TGF- Beta3 (Ttansforming Growth Factor–Beta3 0.5mgTGF- Beta3 (Ttansforming Growth Factor–Beta3)Transfer growth factor - β3(antigen)

NAALADL1Recombinant humanNAALADL1proteinProtein

CDNF Protein HumanRecombinant humanCDNF / ARMETL1protein(Fclabel)

LGALS1 Protein HumanRecombinant humanGalectin-1 / LGALS1protein

After receiving the cells, please proceed with the following steps for the customer. (The number of passages for this cell is limited to about 2. It is recommended that the customer arrange subsequent experiments as soon as they receive the cells.)

1. Remove the cell vial, disinfect it with 75% alcohol, remove the sealing film, and place it in a 37 ℃, 5% CO2 incubator for 6-8 hours to stabilize the cell state.

2. When the cells reach 80% confluence, prepare for subculture.

3. Cell passage:

1) Suck out the culture medium from the cell vial and wash the cells once with PBS.

2) Add about 1mL of 0.125% digestive solution to the culture bottle and digest at 37 ℃ for about 3 minutes; Under the microscope, after the cells shrink and become round, discard the digestion solution and add culture medium to terminate digestion.

3) Gently blow and mix with a straw, inoculate and passage in appropriate ratios such as 1:2 or 1:3, then add fresh culture medium to 5mL and culture in a 37 ℃ 5% CO2 cell incubator.

4) After the cells adhere to the wall, culture and observe, and replace with fresh culture medium every 2-3 days.