GB1027551A - Production of physiologically active placental substances - Google Patents

Production of physiologically active placental substances

Info

Publication number
GB1027551A
GB1027551A GB17497/63A GB1749763A GB1027551A GB 1027551 A GB1027551 A GB 1027551A GB 17497/63 A GB17497/63 A GB 17497/63A GB 1749763 A GB1749763 A GB 1749763A GB 1027551 A GB1027551 A GB 1027551A
Authority
GB
United Kingdom
Prior art keywords
ether
culture medium
acetic acid
tissue
sodium
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired
Application number
GB17497/63A
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
SANFAR LAB Inc
Original Assignee
SANFAR LAB Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by SANFAR LAB Inc filed Critical SANFAR LAB Inc
Publication of GB1027551A publication Critical patent/GB1027551A/en
Expired legal-status Critical Current

Links

Classifications

    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/48Reproductive organs
    • A61K35/50Placenta; Placental stem cells; Amniotic fluid; Amnion; Amniotic stem cells

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Cell Biology (AREA)
  • Developmental Biology & Embryology (AREA)
  • Pregnancy & Childbirth (AREA)
  • Reproductive Health (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • Immunology (AREA)
  • Virology (AREA)
  • Zoology (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

Physiologically active substances such as gonadatropin and steroid hormones are obtained by contacting viable placental tissue with a culture medium in tissue culture apparatus while maintaining the tissue at a temperature suitable to maintain viability and the pH of the culture medium in a viable range, for sufficient time to produce the active substances in the medium and the active substances are then extracted from the culture medium. Trophoblastic tissue in strips 2 inch wide and 6 inch long or, alternatively, a whole intact placenta disposed on a plate with the umbilical cord depending downwardly through an orifice in the plate, is cultured at a temperature of 37 \sB 0.5 DEG C. in a specified culture medium at a pH in the range of 7.2 to 7.8 within apparatus, as depicted in Figs. 1 to 3 (not shown), comprising a cylindrical body defining a closed chamber, viable tissue support means disposed within the chamber, means for intermittently introducing and withdrawing culture medium and means for maintaining the temperature within the closed chamber at a level suitable to maintain viability of a tissue supported therein. The culture medium is stirred by a coated magnetic stirring bar 28 or 56. Semi-permeable membranes of reconstituted cellulose fitted in orifices 101 attached to the walls of the chamber permit the exchange of metabolic products, nutrients and pH-adjusting materials, such as sodium bicarbonate, glutamine or phosphate buffer (as the case may be). Sterile air is circulated through the apparatus. The withdrawn culture medium is cooled down to about 5 DEG C., is acidified to about pH 3.5 with glacial acetic acid and is stored at about 5 DEG C. for 48 hours to precipitate the glycoprotein. The precipitated glycoprotein is vacuum filtered from the medium utilizing activated alumina and a filter aid such as diatomaceous earth, the filter bed being washed with distilled water acidified with glacial acetic acid until the proteinaceious content of the effluent will no longer furnish a precipitate with acetone. The filter cake is thoroughly dispersed in or homogenised with 2N ammonium hydroxide solution and filtered again. The gonadatropic fraction is recovered from the ammonia filtrate by acidification to pH 4.5 with glacial acetic acid and re-precipitation with acetone. The solution is stored at about 5 DEG C. for 48 hours to assure complete precipitation and then the mixture is centrifuged. The precipitate is again dissolved in 2N ammonium hydroxide solution, re-precipitated by addition of acetic acid and acetone, and re-centrifuged. After the removal of residual salts e.g. sodium by dialysis, standardized dosages of the gonadatropin are made up with lactose sodium, hydrogen phosphate (buffer) and freeze dried. Steroid hormones are obtained from the effluent filtrate solution by treating it with diethyl ether, the ether extract is contacted with alcoholic potassium hydroxide to saponify the various fatty acid esters present and soaps are then removed by repeated extraction with distilled water. The ether extract material is then fractionated by passage through a 97%-activated alumina chromatographic column utilizing an elutriant comprising 3% ether in cyclohexane. The separated components are collected as solids by evaporating the elutriant. Alternatively, the saponification purification procedure may be replaced by a chromtographic separation wherein the steroids are absorbed on a 100%-activated alumina column, impurities including fatty acid esters being removed by washing the adsorbate with 100% cyclohexane. The ether-extracted filtrate containing alkaline phosphatase, ribonuclease, adenosine triphosphate, choline esterase, desoxyribonuclease and unidentified components, may be subjected to vacuum to remove residual ether, neutralized to pH 7.8 by sodium hydroxide, and then freeze-dried.ALSO:Physiologically active substances such as chorionic gonadatropin and steroid hormones are extracted from a culture medium at pH 7.2-7.8 in which viable placental tissue has been cultured at a temperature of 37 \sB 0.5 DEG C. by cooling the culture medium to about 5 DEG C., acidifying to about pH 3.5 with glacial acetic acid and storing the medium at about 5 DEG C. for 48 hours to precipitate the glycoprotein which is removed by filtration. The filter cake is thoroughly dispersed in or homogenized with 2N ammonium hydroxide solution and filtered again. The gonadatropic fraction is recovered from the ammonia filtrate by acidification to pH 4.5 with glacial acetic acid and re-precipitation with acetone. The precipitate is separated and again dissolved in 2N ammonium hydroxide solution, reprecipitated by addition of acetic acid and acetone, and re-centrifuged. After the removal of residual salts e.g. sodium by dialysis, standardized dosages of the gonadatropin are made up with lactose and sodium hydrogen phosphate (buffer) and freeze dried. Steroid hormones are obtained from the effluent filtrate solution by treating it with diethyl ether, the ether extract is contacted with alcoholic potassium hydroxide to saponify the various fatty acid esters present and soaps are then removed by repeated extraction with distilled water. The ether extract material is then fractionated by passage through a 97%-activated alumina chromatographic column utilizing an elutriant comprising 3% ether in cyclohexane. The separated components are collected as solids by evaporating the elutriant. Alternatively the saponification purification procedure may be replaced by a chromatographic separation wherein the steroids are absorbed on a 100%-activated alumina column, impurities including fatty acid esters being removed by washing the adsorbate with 100% cyclohexane. The ether-extracted filtrate containing alkaline phosphatase, ribonuclease, adenosine triphosphate, choline esterase, desoxyribonuclease and unidentified components, may be subjected to vacuum to remove residual ether, neutralized to pH 7.8 by sodium hydroxide and then freeze-dried.
GB17497/63A 1962-05-08 1963-05-03 Production of physiologically active placental substances Expired GB1027551A (en)

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
US19315262A 1962-05-08 1962-05-08

Publications (1)

Publication Number Publication Date
GB1027551A true GB1027551A (en) 1966-04-27

Family

ID=22712450

Family Applications (1)

Application Number Title Priority Date Filing Date
GB17497/63A Expired GB1027551A (en) 1962-05-08 1963-05-03 Production of physiologically active placental substances

Country Status (3)

Country Link
CH (1) CH479701A (en)
DK (1) DK108176C (en)
GB (1) GB1027551A (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1025891A1 (en) * 1998-06-17 2000-08-09 Toshia Hanya Method and equipment for treating untreated liquid substance

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1025891A1 (en) * 1998-06-17 2000-08-09 Toshia Hanya Method and equipment for treating untreated liquid substance
EP1025891A4 (en) * 1998-06-17 2001-05-16 Hanya Toshia Method and equipment for treating untreated liquid substance

Also Published As

Publication number Publication date
DK108176C (en) 1967-10-02
CH479701A (en) 1969-10-15

Similar Documents

Publication Publication Date Title
US2301787A (en) Recovery of saponin
US2162863A (en) Stable products shortening the bleeding period
GB1027551A (en) Production of physiologically active placental substances
US2686752A (en) Enzymatic hydrolysis of steroidal saponins
US1758616A (en) Method for recovering quebrachitol from rubber latex serum
US2960500A (en) Process for the extraction and purification of sapogenins
US2190248A (en) Process for obtaining a substance lowering the blood pressure
US2068623A (en) Colored compound derived from yellow oxidation ferment and method of producing same
US3170839A (en) Culture process in vitro placental tissue growth medium
ATE70452T1 (en) PROCESS FOR PRODUCTION OF A PROPOLIS|THANO EXTRACT.
DE839643C (en) Process for the purification of p-aminosalicylic acid
US1976175A (en) Process of preparing adenosine phosphoric acid
US3442916A (en) Isolation of eburicoic acid
SU568676A1 (en) Source of raw material for obtaining ergosterine
US2837540A (en) Extraction of sterols
US3530114A (en) Isolation procedures of coumermycin a1
DE577258C (en) Process for the production of hormones regulating cell growth
JPS61180719A (en) Method for obtaining ganglioside
DE679538C (en) Process for the production of a product influencing oral and jaw diseases
GB479487A (en) Manufacture of stable therapeutic products from intestinal mucous membrane
US1902408A (en) Manufacture of new cardio-active glucosides of bulbus scille
US2744892A (en) Crystallization of dihydrostreptomycin sulphate
US2799618A (en) Glycosidal material having activity on the heart and vascular vessels and its preparation from plant sources
US2027446A (en) Process for preparing gonad stimulating hormone
US730197A (en) Process of isolating the active principle of the suprarenal glands.